Latex Test Research Articles

Information value of latex test for pathogenic burkholderia serological diagnostics using microscopy and video digital recording system

Objective: the purpose of the work is assessment of the possibility of using instrumental assay to record the results of the latex test in the detection of pathogenic burkholderia. Monoclonal based latex agglutination test was prepared according to a previously developed technique. The latex agglutination test results were obtained using a “Nikon Eclipse TE 2000-U HMC/System microscope”, as w ell as the video-digital hardware-software com plex “Expert-lab”. The results of an objective assessment of the latex agglutination test were obtained using statistical analysis. The use of “Nikon Eclipse” microscope for visual accounting of microagglutination increased the sensitivity of the latex test to 1•107 CFU/ml. The “Expert-lab” make arrangements for quantitative assessment of the scanned image of the latex test results, which makes it possible to determine the threshold values for discrimination between positive and negative samples. Objective information about the presence or absence of agglutination is recorded. Numerical values of the latex test intensity between samples and controls are compared using system software. Also we can a possibility to save protocols and images of the study, to archive the obtained data. The use of “Nikon Eclipse TE 2000-U HMC/System” microscope for visual recording of the results and on the “Expert-lab” video digital system for objective evaluation of data when searching for pathogenic burkholderia allow s us to document and to archive the data obtained, to conduct in-laboratory quality control of studies and increase the sensitivity of the latex-agglutination reaction.

Assessment and correction of local antimicrobial protection factors in women with chronic recurrent vulvovaginal candidiasis and type 2 diabetes mellitus

Aim. Evaluation and correction of local antimicrobial protection factors in women with type 2 diabetes mellitus (DM 2) and chronic recurrent vulvovaginal candidiasis (rVVC). Materials and methods. The study involved 100 women undergoing outpatient follow-up at the age of 40.9±5.8 years, with a body mass index of 29.8±3.5. The period was 2022–2023. The examination included anamnesis, collection of anthropometric data, calculation of body weight, microscopic examination of smears from the cervical canal and vagina, Gram-stained before treatment, in the 1st and 3rd months after its completion, identification pathogenic microorganisms using PCR, colposcopy, ultrasound of the pelvic organs. Indicators of glycosylated hemoglobin in the subjects at the control points of the study: at 1 month the average values were 5.9±2.9%, at 3 months – 5.9±3.0%, the average value was 5.9±3.1%. Candida species were identified by the bacteriological method using Sabouraud dextrose agar (growth of colonies of fungi of the genus Candida in an amount of more than 103 CFU/ml). Immunological methods for studying antimicrobial protection factors included studying the number of neutrophil granulocytes (NG) on the surface of the mucous membranes of the vulva and vagina, their phagocytic and NBT-reducing activity in a latex test. Randomization of patients into groups: group 1 included 50 (50%) women with DM 2 who, as part of complex therapy for rVVC received the drug Genferon® 1 intravaginal suppository of 500 thousand units 2 times a day for 10 days and fluconazole 150 mg orally three times with an interval of 72 hours at the first stage of treatment; group 2 included 50 (50%) women who received therapy with fluconazole 150 mg three times with an interval of 72 hours. The maintenance anti-relapse course therapy in both groups included the use of fluconazole 150 mg once a week for 6 months. In group 1st anti-relapse therapy was supplemented by the administration of the drug Genferon® 1 intravaginal suppository of 500 thousand units at night 3 times a week for 3 months, after which the vaginal microbiota was corrected for 2 weeks using vaginal suppositories, containing Lactobacillus acidophilus in an amount of at least 108 CFU. In group 2nd, basic anti-relapse antimycotic therapy was not accompanied by the prescription of any immunomodulatory or probiotic drugs. Results. The etiological agents of rVVC in women of late reproductive age in 79% (n=79) и 28% (n=28), respectively, were C. albicans and C. glabrata, which during the period of acute of the disease were detected by culture in the vaginal discharge at more than 104 CFU/ml. Chronic recurrent course of vulvovaginal candidiasis in women with DM 2 was characterized by exacerbations 4 or more times a year, accompanied by the corresponding clinical picture: white or yellowish-white cheesy discharge from the genital tract, itching or burning in the anogenital area, discomfort in the external genital area, dyspareunia, dysuria, decreased phagocytic activity of NG in vaginal secretions by 25.8%, impairment of their spontaneous and latex-induced NBT-reducing activity by 35.2%, functional reserve by 23.0% relative to reference values. The use of the drug Genferon® as part of complex therapy for rVVC contributed to a decrease in the number of Candida spp. in 1st and 3d months of observation after completion of anti-relapse therapy, normalization of cellular factors of innate immunity of the mucous membranes, faster resolution of the clinical manifestations of an episode of the disease. The decrease in the number of relapses over a 12-month observation period compared to the control group was also facilitated by an increase in the protective properties of the vaginal mucosa due to the restoration of the vaginal microbiota with the help of lactobacilli acidophilus. Conclusion. The etiological agents of rVVC in women of late reproductive age are C. albicans and C. glabrata. Subcompensated DM 2 is a risk factor for rVVC, which requires additional monitoring of microbiological parameters of the vaginal microbiota. rVVC in women with DM 2 is associated with an increase in the number of NG in the vaginal secretion, a decrease in their phagocytic activity and functional reserve compared to the reference values. Combination therapy of rVVC with topical recombinant interferon α2b, benzocaine and taurine in the formulation of Genferon® (suppositories) is an effective method to improve the clinical and immunological efficacy of therapy.

Investigating the Occurrence of <i>Escherichia coli</i> O157:H7 in the Final Effluents of Two Wastewater Treatment Plant in the Eastern Cape of South Africa

The final effluents of two wastewater plants located in the Eastern Cape of South Africa were tested for the presence of enterohaemorrhagic Escherichia coli O157:H7 isolates, and characteristics of the isolates obtained were determined. A total of 23 wastewater samples were collected from the treatment plants at the final effluent point after the disinfectant stages of wastewater processing. Altogether, 540 presumptive E. coli isolates were obtained by colony counting on the E. coli O157:H7 chromogenic agar base supplemented with cefixime tellurite and were subcultured onto sorbitol-MacConkey agar and tested for agglutination using the Prolex E. coli O157 latex test reagent kit.The results showed that the 149 suspected colonies from SMAC agar were all negative for the antisera. None of the isolates agglutinated with antisera against E. coli O157 and thus no presence of the bacteria can be confirmed from the treated effluents.The likelihood of the receiving water body and the environment being contaminated with E. coli O157:H7 is therefore minimal. Future monitoring is however recommended

Yeasts as a Potential Causes of Surgical Site Infections at the Sourô Sanou University Hospital Center of Bobo-Dioulasso, Burkina Faso

Fungal surgical site infections (SSIs) trend to increase in the recent decades. This study aimed at estimating the prevalence of fungal SSIs, and the time of occurrence of fungal SSIs at the Sourô Sanou University Hospital (SSUHC) in Burkina Faso. This study was carried out in two departments of the SSUHC. Samples of postoperative purulent secretions were collected from June to December 2019. Purulent secretions swabs at the surgical site were aseptically collected using sterile cotton swabs. Standard mycology, CHROMagar, agglutination latex test and immunological test were used to identify the different Candida species. A total of 39 postoperative patients were included in the study. The median age was 27 years and the majority of patients were women (74.4%). The probability of isolating fungal agents from the surgical site was 2.6% three days after surgery. This probability was 14.4% and 49.5% at one week and 12 days after the surgical intervention, respectively. A total of 8 (20.51%) fungal species, all belonging to the genus Candida were identified in the study. These species were Candida albicans (3), Nakaseomyces glabrata (2), Candida tropicalis (2) and Pichia kudriavzevii (1). This study showed that Candida spp. could be the cause of the occurrence of SSIs at the SSUHC in Burkina Faso. So, physician must integrate the fungal causes in the management of SSIs.

Prevalence of Esbl-Producing Enterobacteriaceae Strains Isolated to the University Hospital Center of N’djamena and their Sensitivity to Antibiotics

Introduction: The resistance of Enterobacteriaceae to antibiotics is experiencing a worrying worldwide evolution with a growing impact in the community and medical sector. The difficulties encountered in hospitals in treating certain infections resulting from multi-resistant bacteria are a source of great concern in human health. The aim of this study was to determine the prevalence, diversity and antibiotic resistance of Extended-Spectrum Beta-Lactamase (ESBL)-producing Enterobacteriaceae in the hospital setting. Materials and Methods: A total of 830 specimens from hospitalized and outpatients including 341 stools, 257 urine and 232 pus were collected between August 2017 and February 2019. Enterobacteriaceae were identified using the Api 20E gallery and the VITEK® 2TM 15 compact automaton. E. coli and Salmonella serotyping were performed by serum agglutination using the Latex Test Kil and the Kauffmann-White Kit, respectively. Sensitivity tests according to the Antibiogram Committee of the French Society of Microbiology and double disc synergy to detect ESBLs have been carried out. Results: Out of the 830 specimens, 181 (21%) strains were isolated at the N’Djamena National Reference University Hospital between August 2017 and February 2019. 56 strains produced ESBLs, i.e. a prevalence of 30.9% (56/181). Escherichia coli was more common (74%), followed by Klebsiella pneumoniae (6.6%), Proteus spp (14.9%), Salmonella (2.8%) and Shigella (1.7%) (p<0,05). These germs were isolated from urine at 47%, pus at 38% and stool at 15%. Resistance with E. coli was respectively 60.8% and 50% for imipenem and amoxicillin + clavulanic acid. Regarding Klebsiella pneumoniae, these rates were 70% and 55% respectively. Salmonella showed resistance to ciprofloxacin (80%), gentamicin (80%) and ceftriaxone (60%). E. coli, Klebsiella pneumoniae have been the most implicated with strong resistance to the majority of antibiotics. Conclusion: This study highlighted the diversity of enterobacteria causing diarrhea and the emergence of multi-resistant bacteria. It helps to update knowledge for better patient care.

Rheumatoid factor isotypes in rheumatoid arthritis diagnosis and prognosis: a systematic review and meta-analysis

ObjectiveThe first biomarker associated with the rheumatoid arthritis is rheumatoid factor (RF) and since the earliest reports a role has been proposed in the diagnosis and in the prediction of...

Microbiological Evaluation of Different Types of Branded and Non-branded Ready-to-Eat Snacks Sold in Elementary Schools of District Peshawar, Pakistan

The lack of hygiene in the preparation and packing of ready-to-eat foods is fast becoming a serious public health concern, especially for school children. The intervention of different pathogenic microorganisms in these foods poses the risk of foodborne disease outbreaks. The current study was designed to assess bacterial contamination in different types of ready-to-eat (branded and non-branded) snacks purchased from September 2021 to December 2021 in various elementary schools of Peshawar, Pakistan. A total of 20 samples were collected and analyzed using the pour plate method for total plate count (TPC). Moreover, the multiple fermentation tube method was used for total coliforms (TC) and fecal coliforms (FC). Escherichia coli isolates were identified using E. coli O157:H7 latex test reagent kit Pro Lab, Canada. The results revealed contamination by TPC (6.67%), TC (40%), FC (33.33%), and E. coli (20%) in the samples. These values were higher than the permissible limits set by Food and Agriculture Organization (FAO). Hence, it was concluded that there is a practice gap in food safety knowledge among ready-to-eat food vendors. The vendors are usually untrained and lack the knowledge of proper hygiene and food handling procedures. It is suggested here that the government should pay special attention towards improving public awareness regarding food safety and quality of ready-to-eat foods sold in Peshawar, Pakistan.

Toxoplasmosis in Females from Al-Anbar, Iraq.

الخلفية: داء المقوسات هو مرض حيواني المنشأ شائع يصيب ثلث سكان العالم مع آثار صحية خطيرة محتملة. هدف الدراسة: تحديد مدى انتشار وعوامل الخطورة للإصابة بمرض التوكسوبلازما بين النساء في محافظة الأنبار، العراق. المرضى والطرق: على مدى 8 أشهر من الأول من تشرين الثاني 2021 إلى 30 يونيو 2022، شاركت 200 سيدة في زيارة عيادات بيطرية خاصة في الأنبار (92 منهن كان لديهن قطط) في هذه الدراسة المستقبلية. تم تسجيل عمر ومكان اقامة النساء المشاركات في الدراسة. تم جمع عينات الدم من جميع النساء وخضعت العينات أولاً لاختبارتلازن اللاتكس السريع (TLAT). جمعت عينات البراز من 57 (62٪) قطة أليفة و 35 (38٪) قطة ضالة وخضعت عينات البراز أولاً للفحص المجهري المباشر بحثاً عن بيوض التوكسوبلازما. ثم تم اختبار عينات الدم والبراز عن طريق تفاعل البلمرة المتسلسل (n-PCR) وتفاعل البلمرة المتسلسل في الوقت الحقيقي (RT-PCR) ثم تم تحليل النتائج إحصائيا. النتائج: كانت معدلات داء المقوسات البشري (ن = 48، 24٪)، (ن=60، 30٪)، و (ن= 77، 38.5٪) باختباراتTLAT ، n-PCR ، و RT-PCR على التوالي. وبالمثل، كانت معدلات داء المقوسات في القطط (ن = 7، 7.61٪)، (ن = 55، 60٪)، و (ن = 80، 87.0٪) باختباراتTLAT ،n-PCR ، و RT-PCR على التوالي. أظهرت نتائج اختبار اللاتكس أن النساء فوق ال 40 سنة كان لديهن معدل إصابة مرتفع (45.8٪) مقارنة باللواتي تتراوح أعمارهن بين 31-40 سنة (33.8٪) واللواني تتراوح أعمارهن بين 21-30 سنة (13.0٪ .(P≤0.01) (كان معدل إصابة النساء في المدينة أقل مقارنة بالقرية (29.7٪ مقابل 55.4٪). الاستنتاجات: وجدنا من خلال هذه الدراسة ان داء المقوسات أكثر انتشارًا بين النساء فوق ال 40 سنة وفي المناطق الريفية ولكنه لم يكن مرتبطًا بملامسة القطط. وفر تفاعل البلمرة المتسلسل في الوقت الحقيقي (RT-PCR ) عائدًا تشخيصيًا أفضل. حتى يتم اكتشاف لقاح، تظل النظافة أفضل إجراء وقائي لداء المقوسات البشري.

Evaluation of an on-farm culture system for the detection of subclinical mastitis pathogens in dairy cattle

The purpose of this observational study was to compare the performance of a novel on-farm culture (OFC) test with the reference method (RM) in identifying pathogens, and in particular Staphylococcus aureus, associated with subclinical mastitis (SCM) in dairy cattle. The OFC test (Mastatest HiSCC; Mastaplex Limited) for SCM uses a cartridge with 2 × 12 wells allowing 1 sample to be analyzed in duplicate (24 wells) or 2 samples analyzed simultaneously, each in 12 wells. Results of the milk analyses are reported hierarchically (Staph. aureus → coagulase-negative staphylococci (CNS) → other gram positive or coliform/gram negative → no bacteria present) and emailed within 24 h. Milk samples (617 quarter level from 158 cows and 70 cow level) were collected from 288 cows [individual cow somatic cell count (ICSCC) ≥150,000 cells/mL] on 9 purposefully selected farms known to have a high prevalence of clinical and subclinical Staph. aureus mastitis in Southland New Zealand. Quarter samples were analyzed individually (617 samples) and after animal-level pooling, providing 228 (158 + 70) cow-level samples. Samples were analyzed by the OFC test (in duplicate) and the RM (culture agar medium and latex test based on the recommendation by the National Mastitis Council) and identifications confirmed with MALDI-TOF mass spectrometry. Sensitivity (Se), specificity (Sp), positive predictive value (PPV), and negative predictive value (NPV) for detection of Staph. aureus were all ∼90% with tight 95% confidence limits, and Cohen's kappa (κ) for agreement between the OFC test and RM was 0.81. Kappa for agreement between the OFC test duplicates was 0.93. About 35% of cows had only one quarter infected with Staph. aureus and all these animals could still be identified when pooled cow-level milk was analyzed. Although the high prevalence of Staph. aureus in the herds used in this study does not affect the Se and Sp values, it does elevate the PPV value (and decrease the NPV) and therefore use of PPV to extrapolate to a population with lower prevalence is not appropriate. For CNS, Sp, PPV, and NPV were all >0.8, κ was ≥0.6, and Se was >0.7. Kappa for agreement between the OFC test duplicates was 0.83. A result of “no bacteria detected” was reported in 13% of the cows with 93% agreement between OFC test and RM. We conclude that the OFC test provides a reliable method for detecting Staph. aureus in pooled cow-level milk even if only one quarter is infected; in the absence of Staph. aureus in the milk, it reliably identified CNS in pooled cow-level milk; it reliably identified cows with <10 cfu/10 µL of their milk. Compared with the RM, the method was rapid with results returned in 24 h of loading the cartridge.

Detection of Legionella pneumophila in the Water Samples of Food Industries and Hospitals in Bangladesh

Background: The human pathogen Legionella pneumophila causes a serious pneumonia-like respiratory disease called Legionnaires' disease, mainly in elderly and immunocompromised individuals. This pathogen can be found in the water distribution systems of large constructions with cooling towers which is a common phenomenon at present in Bangladesh due to its rapid economic growth. But there is a dearth of information on the incidence of Legionella in Bangladesh. Therefore, the current study aimed to investigate the presence of Legionella pneumophila in hospital and industrial water distribution systems in Dhaka, Bangladesh. Methods: A total of 114 water samples collected from two hospitals and five food industries were inoculated on the Legionella-specific medium Buffer Charcoal Yeast Extract (BCYE) agar medium before and after the treatment with acid, heat, or a combination of both. Samples producing Legionella-like colonies on BCYE agar medium were screened by Legionella Latex Test Kit, and the metagenomic DNAs obtained from these samples were analyzed by PCR using L. pneumophila-specific 16S rRNA primers. Results: Among 114 samples, Legionella-like colonies were observed in 30 water samples which demonstrated no agglutination in the Latex agglutination test. PCR analysis showed the presence of L. pneumophila in seven water samples, four in the potable water, chiller water, and cooling tower water of two different food industries, and three in ICU tap water, cooling tower water, and Fan Coil Units of two different hospitals. Sequence analysis of amplicons revealed that all seven sequences had 100% similarity with L. pneumophila. Conclusion: The presence of L. pneumophila in the water samples of local hospitals and food industries indicates that these habitats might serve as a potential site for Legionnaires' infection in Bangladesh. The results also showed that PCR, contrary to the conventional culture methods, could be more efficient and rapid in the identification of L. pneumophila.

Cold-Induced Urticaria Exacerbated During Treatment With Pembrolizumab

An 80-year-old woman was assessed for cutaneous edema at the site of her intravenous (IV) access during infusion of normal saline (Figure 1). She had no associated symptoms and was otherwise well. The same reaction occurred in the contralateral arm with dextrose 5%. She was treated with IV diphenhydramine and the reaction subsided. The patient was on a research protocol including cyclophosphamide, pembrolizumab, and antisurvivin tumor vaccine to treat refractory diffuse large B-cell lymphoma. Her previous medical conditions included chronic hepatitis B on tenofovir and hypothyroidism on levothyroxine. She described a more than 20-year history of cold intolerance, reporting local hives with exposure to subzero degree centigrade temperatures, as well as throat tightness and epigastric pain after consuming frozen foods. She managed her symptoms by cold avoidance alone. Her symptom severity increased after starting pembrolizumab. Cold-induced urticaria (CIndU) was suspected on the basis of history and hypothesized as the cause of her cutaneous reaction to room temperature saline. To support the diagnosis of CIndU, skin testing was performed. Latex and chlorhexidine skin prick test (SPT) results were negative, ruling out the possibility of contact urticaria. Results for the ice cube test and SPT with room temperature saline were both positive (60 mm and 40 mm wheals, respectively), with histamine control of 4 mm; SPT result to saline warmed to 30°C was negative. After pretreatment with 20 mg of cetirizine, repeat SPT result to room temperature saline was negative. She then tolerated challenge to an IV infusion of 250 mL of 30°C saline. In the context of her history and diagnostic testing, CIndU triggered by room temperature IV fluid was diagnosed. The patient was started on daily oral antihistamine therapy (20 mg of bilastine). Subsequent IV infusions were successfully administered by heating the IV lines at the point of insertion to 30°C. She was able to receive additional chemotherapy doses in this manner but was moved to palliative care before any additional testing could be performed. Interestingly, although there is often no identified underlying disease trigger for CIndU, the comorbidities of our patient (hypothyroidism, chronic hepatitis B, malignancy) are all established potential etiologies. However, her CIndU predated these health conditions and did not worsen until pembrolizumab therapy. The cold protein diseases (cryoglobulins, cryofibrinogen, or cold agglutinins) should also be considered in the differential of cold-exacerbated diseases, especially cryoglobulinemic vasculitis, which can be linked to chronic hepatitis and malignancy. However, cold proteins have not been clearly linked to the pathophysiology of CIndU, and this patient was not experiencing symptoms compatible with vasculitis or worsening anemia, making cold protein disease unlikely.1Maltseva N. Borzova E. Fomina D. Bizjak M. Terhorst-Molawi D. Košnik M. et al.Cold urticaria – what we know and what we do not know.Allergy. 2020; 76: 1077-1094Crossref PubMed Scopus (30) Google Scholar Why was the patient’s CIndU acutely exacerbated after many years of stable disease? We hypothesized that pembrolizumab might have lowered the patient’s CIndU temperature threshold. Indeed, pembrolizumab is an IgG4 antibody that inhibits the programmed cell death protein 1 (PDI) inhibitor on lymphocytes. PDI is an “immune checkpoint” on activated effector T and B cells that binds to its cognate receptor PD-L1, inhibiting activated lymphocytes. Many cancer cells produce PD-L1 as an immune evasion strategy. Pembrolizumab removes this inhibition, enabling lymphocytes to better target and attack cancer cells. However, pembrolizumab also has many off-target effects, mainly immune-mediated adverse events. Cutaneous adverse events (CAEs) are the most common type of adverse event occurring on pembrolizumab, affecting approximately 25% of patients. Most CAEs are nonspecific rash or pruritis, but urticaria can occur in 0.7% of patients.2Wongvibulsin S. Pahalyants V. Kalinich M. Murphy W. Yu K. Wang F. et al.Epidemiology and risk factors for the development of cutaneous toxicities in patients treated with immune-checkpoint inhibitors: a United States population-level analysis.J Am Acad Dermatol. 2022; 86: 563-572Abstract Full Text Full Text PDF PubMed Scopus (22) Google Scholar A mechanism for exacerbating preexisting CIndU is not established. Extrapolating from other immune-mediated CAEs on pembrolizumab, such as bullous pemphigoid and vitiligo, PD-1 blockade may have caused increased autoantibody formation (IgE or IgG autoantibodies to FcεRI or IgE), thus decreasing the patient’s CIndU threshold. Chronic urticaria index or autologous skin testing, when positive, would have provided evidence of autoantibody formation to support this hypothesis, but was not possible for this patient. This case highlights the need for ongoing research on both the pathophysiology of CIndU and the expanding field of immune-mediated side effects of checkpoint inhibitors.

Frequency of infectious disease caused by Methicillin sensitive Staphylococcus aureus (MSSA) among community and hospital acquired infections in Khartoum State, Sudan

Background: S. aureus is the most common bacteria associated with both community and hospital associated infections. It is associated with skin and wound infection, pneumonia, endocarditis, septic arthritis, osteomyelitis. Objectives: The present study aimed to estimate the prevalence of infectious disease caused by Methicillin sensitive Staphylococcus aureus among community and hospital acquired infections in Khartoum State, Sudan. Methods: A cross-sectional laboratory-based study was conducted during the period from November 2020 to January 2021. To isolate and identify S. aureus, the conventional method and protein A latex test were used. All isolated organisms were tested for their in-vitro antimicrobial susceptibility against methicillin (MET 5μg) disc using the Kirby-Baur disk diffusion method to detect MSSA. Result: The result showed a high frequency of community infection (80%). The majority of the isolates were isolated from patients with respiratory tract infection (28.7%), and the result showed that RTI more common on community infection (33.7%). Conclusion: The present study concluded that male more infected by MSSA and the highest frequency of isolates was in the age group more than 45 years. The present study highlighted that RTI is predominantly the site of infection by MSSA. The present study showed that RTI more common on community infection.

Methicillin-Resistant Staphylococcus aureus in Diabetic Foot Infections: Protein Profiling, Virulence Determinants, and Antimicrobial Resistance

Staphylococcus aureus (S. aureus) is one of the most prevalent bacterial pathogens recovered from diabetic foot infections (DFIs). Most S. aureus isolates exhibit methicillin resistance, so treatment is recommended with antimicrobials active against methicillin-resistant S. aureus (MRSA) in patients who have risk factors associated with MRSA infections. The main goal of this study was to see if proteomics and molecular methods could be effective in identifying and distinguishing MRSA recovered from DFIs. Since MRSA is highly resistant to β-lactam antibiotics and usually does not respond to other antimicrobial drugs, we evaluated the resistance of MRSA isolates against different antibiotics. The standard procedures were followed for a culture of 250 skin swabs collected from diabetic foot patients. The phenotypic characteristics of 48 suspected S. aureus cultures were determined via microscopic examination, Gram staining, a coagulase test, a BBL™ Staphyloslide™ Latex test, a Staph ID 32 API system, and a Vitek 2 Compact system. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to examine the protein profile of all isolates, and real-time PCR was then used to identify mecA and PVL virulence genes. S aureus isolates were tested using the Vitek 2 Compact for antimicrobial susceptibility using Gram-positive cards (GP71). Among the 48 bacterial isolates tested, 45 (93.75%), 42 (87.5%), and 46 (95.83%) were positive in tube coagulase, the Staph ID 32 API system, and the Vitek 2 Compact system, respectively. We correctly identified all suspected S. aureus isolates (100%) via MALDI-TOF MS with a score value ≥2.00 and differentiated them into 22/48 MRSA (45.83%) and 26/48 MSSA (54.17%) isolates. A higher peak intensity at masses of 5530 Da, 6580 Da, 6710 Da, and 6820 Da was detected in MRSA, but not in MSSA. All MRSA isolates tested positive for the mecA gene, while all isolates tested negative for the PVL gene. The antibiotic susceptibility results showed that 22 (100%), 20 (90.91%), 19 (86.36%), 18 (81.82%), 17 (77.27%), 15 (68.18%), 13 (59.1%), and 12 (54.55%) MRSA strains were resistant to cefoxitin, daptomycin, erythromycin, benzylpenicillin, ciprofloxacin, oxacillin, and clindamycin, respectively. In contrast, all MRSA strains were extremely susceptible (100%) to linezolid, nitrofurantoin, quinupristin–dalfopristin, tigecycline, and vancomycin. Moreover, 20 (90.91%), 18 (81.82%), and 17 (77.27%) of the MRSA strains exhibited high sensitivity against rifampin, trimethoprim–sulfamethoxazole, and gentamicin, respectively. In DFIs, MALDI-TOF MS is a powerful and accurate method of identifying and distinguishing both MRSA and MSSA isolates. A high level of antimicrobial resistance was found in MRSA isolates, and antibiotic therapy based on antibiotic susceptibility patterns is essential for a successful outcome.

Microscopic and ELISA Based Detection of Toxoplasma and its Associated Risk Factors in Pregnant Women of District Mardan

Toxoplasmosis caused by the protozoan parasite Toxoplasma gondii, the most prevalent disease worldwide. The infections in human are extremely dangerous if they happen during pregnancy because they may lead to miscarriage or congenital abnormalities, which predominantly affect the central nervous system. The main objective of current study was to identify the risk factors for toxoplasmosis and its prevalence among pregnant women in Mardan. The blood serum was tested using the ELISA and the latex agglutination test. Through interviews, sociodemographic and prospective risk factor data were gathered using a standardized questionnaire. Our findings, obtained from ELISA and latex agglutination tests, showed considerably higher toxoplasmosis rates of 23% and 11%, respectively. The geographic distribution of toxoplasma infection in district Mardan revealed a high prevalence in Shah Baig village through ELISA (28%) and latex test (12%) followed by Ibrahim Khan Killi (24%) and (16%), while lower cases were recorded through ELISA (20%) in Sheikh Yousaf Killi and Saleem Khan Killi. Compared to working women in the community (5.85 percent), the infection was more prevalent (18%) in non-occupational women/house wives. Moreover, it was also observed that abortion occurred to the woman who have been clinically diagnosed positive by ELISA and it reported 20% by ELISA and 11.4% by latex agglutination test. It was concluded that T. gondii infection were more prevalent in pregnant women of rural areas of Mardan as compared to the urban centers. Women and children were more vulnerable if they frequently interact with domestic animals and play in contaminated environment.

SAPHO Syndrome: About an Observation and Review of the Literature

Introduction: SAPHO (synovitis, acne, pustulosis, hyperostosis, and osteitis) is autoinflammatory rheumatism of the spondyloarthritis group. It is rare, it prevalence is 1/10000 inhabitants. The average age at diagnosis is 38 years, with no gender predominance, first described by Chamot and colleagues in 1987. Objective: To present observation and review of the literature on SAPHO syndrome which is rarely diagnosed in our daily practice. Observation: This was a 28 year old female patient, Senegalese, living in Dakar, followed in obstetrics gynecology for the desire of maternity, the family context of sickle cell disease who consulted for diffuse mechanical and articular inflammatory pains involving the proximal interphalangeal (IPP) of the left 5th toe, the left shoulder, the sternocostal and dorsolumbar joints evolving for 3 years. The clinical examination found: A painful swelling with a visual analog scale (VAS) of 6/10 of the right sternocostal joint and of the interphalangeal joint (IP) of the right hallux, diffuse acne, no palmoplantar pustulosis, no psoriasis, no gluteal or talalgia. The biology shows an inflammatory syndrome (VS at 37 mn at the 1st hour and CRP at 41mg/l), a decrease of 25-OH-Vitamin D3 at 22 ng/ml (N: 25-30 ng/ml), the positive latex test, Ac anti-CCP: 43 IU/ml (N &lt; 5 IU/ml), positive Ac anti-HBc , Ac anti HBs &gt; 1000 IU/l on the other hand blood calcium, blood phosphorus, G6PD, PTH, TSH us, free T4, anti ECT Ac, HLA-B27 Ag with no particularities. The scintigraphy showed hyper fixation of the right 1st toe, sacroiliac joints, dorsolumbar spine, left shoulder, manubrium sternal junction, and bilateral sternoclavicular junction realizing a classical bull horn image in SAPHO syndrome. Because of this clinical and paraclinical manifestation helped by the 1994 diagnostic criteria of Kahn at 1/3 (chronic joint involvement associated with severe acne), the SAPHO syndrome was retained and the treatment instituted after the normal pre-therapeutic analysis and systematic deworming was: Salazopyrine 2000mg/day (dosage reached by increments of 500 mg/week); Uvedose (vitamin D3 ) 100000 IU / month; bisphosphonates 70 mg /week and paracetamol + codeine 1g x 2/day as needed for 3 months with a favorable follow-up by the regression of signs (pain VAS at 2/10). Conclusion: The SAPHO syndrome is the prerogative of the young adult subject, bone scintigraphy is very valuable for its diagnosis, its better knowledge would avoid diagnostic and therapeutic erratic. Keywords: SAPHO syndrome, bone scan, Dakar, Senegal.

P243 Case series of Cryptococcal Meningitis—Experience in North Western India over 1 year (2021–22)

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMObjectiveCryptococcosis is an opportunistic fungal infection causing high morbidity and mortality in patients, preferentially affecting immunocompromised. It can cause a wide array of clinical manifestation, which includes meningitis, pulmonary, as well as disseminated infection. Cryptococcus neoformans causes more than 90% cases of cryptococcal meningitis.MethodologyWe performed a retrospective review of patients with confirmed cryptococcal meningitis during 1 year period from 2021 to 2022 in tertiary care center, AIIMS Jodhpur. We assessed clinical, radiological, microbiological, and biochemical parameters along with treatment provided and outcomes of the patient.ResultsOf 189 patients screened for suspected cryptococcal meningitis, 6 were microbiologically confirmed positive. All the patients were immunocompromised, of which four were HIV positive and one was a solid organ transplant recipient on immunosuppression and one was old TB Meningitis. Most common symptom was headache and altered sensorium (100%). Radiological findings showed 30% had no significant abnormality. CSF examination revealed average CSF protein 97.6 (63-163), CSF chloride 103.3 (108-132), sugar 36.33 (1-68), with predominant lymphocytes. All the patients were microbiologically confirmed by CSF cryptococcal latex test. A total of 4/5 patients received amphotericin B (3 mg/kg) with fluconazole (1200 mg) for 2 weeks in the induction phase followed by fluconazole consolidation phase and maintenance phase. Of the five patients, four patients survive with a good response to the treatment with one fatality.ConclusionThrough our case series we emphasize the fact that Cryptococcal meningitidis may present with non-significant radiological features. Thus, the differential diagnosis of C. meningitidis must always be thought of when an immunocompromised patient presents with headaches and other signs and symptoms involving the central nervous system.

Viscosity prediction model of natural rubber-modified asphalt at high temperatures

Viscosity is a physical indicator of the flow behavior of asphalt at high temperatures, and the mixing and compaction quality of asphalt mixtures depends on it; however, previous studies cannot effectively guide the application of natural rubber-modified asphalt (NRMA). In this investigation, the effects of the natural rubber latex (NRL) dose, test temperature and shear rate on the apparent viscosity of NRMA were evaluated. The results show that viscosity is approximately positively correlated with NRL dose, as described by an exponential equation that is based on Einstein's equation. Meanwhile, viscosity is negatively correlated with test temperature and shear rate, in accordance with the Arrhenius equation and power law equation, respectively. A three-factor physical equation was established to describe the relationship between viscosity and NRL dose, temperature and shear rate on the basis of the equations above, which can predict the viscosity over a wider range with limited experimental data, thereby overcoming experimental limitations. This study provides a prediction model for quickly determining the viscosity. This model is not only applicable to NRMA but also can provide references for the study of viscosity prediction for other polymer-modified asphalts.

Prevalence of antinuclear antibodies and rheumatoid factor titers in dogs with arthritis secondary to leishmaniosis (Leishmania infantum).

Dogs with infectious arthritis may occasionally exhibit positive serum antinuclear antibody (ANA) and rheumatoid factor (RF) titers; however, relevant data are sparse for arthritis secondary to canine leishmaniosis (CanL) caused by Leishmania infantum. We determined the prevalence of positive serum ANA and RF titers in dogs with arthritis secondary to CanL. Blood samples from adult, client-owned dogs with purulent arthritis secondary to CanL, without any comorbidities, were collected for diagnostic purposes. Serum ANA titers were measured by immunoperoxidase test and RF titers by the Rose-Waaler latex test. Twelve of 23 dogs enrolled prospectively in our study had clinical arthritis, and 11 of 23 had subclinical arthritis. Based on LeishVet clinical staging, 7 dogs had clinical stage II disease, 11 had clinical stage III disease, and 5 had stage IV. None of the 23 dogs was seropositive for ANA; 3 of 23 were positive for RF. ANA and/or RF seropositivity, in dogs with CanL-associated arthritis, appears to be weak, if present at all. Based on our results, positive serum ANA and RF titers should not be expected in dogs with arthritis secondary to CanL.

РОЛЬ ИММУНОЛОГИЧЕСКИХ ФАКТОРОВ В НЕБЛАГОПРИЯТНОМ ИСХОДЕ ХРОНИЧЕСКОЙ ОБСТРУКТИВНОЙ БОЛЕЗНИ ЛЕГКИХ

Aim. The aim of the research was to study the role of immunological mechanisms in the unfavorable outcome of chronic obstructive pulmonary disease (COPD). Material and methods. The research included 116 patients hospitalized in 2005-2006 in the pulmonology department of patients with the exacerbation of mild and moderate COPD. Research protocol: general clinical and special (immunological) methods of examination were performed in patients with COPD. Instrumental research methods included: spirography, ECG, EchoCG, fibrobronchoscopy, chest X-ray. After diagnostic bronchoscopy, bronchoalveolar fluid (BAF) was taken for cytological and immunological examination. On the first and second days of inpatient treatment, patients underwent immunological studies: immunophenotyping of mononuclear cells (MNC), assessment of phagocytic activity of leukocytes (latex test, nitroblue tetrazolium (NBT) test) in blood and BAF, determination of the content of immunoglobulins in blood serum and BAF, study of cytokine levels in blood serum, BAF and MNC culture supernatant under conditions of their spontaneous production and activation in the mitogen-stimulated (phytohemagglutinin "Difco" 5 mg/ml) proliferation. Statistical data processing was carried out using the Statistica 10.0 program. In 2020, the long-term survival of COPD patients was assessed. The cohort of patients was divided into two groups. Survivors were included in the first group (n=44), and patients who died by 2020 were included in the other group (n=72). The retrospective comparison of the studied indicators was determined at the time of COPD exacerbation in 2005- 2006 in these patient groups. Results and discussion. In the surviving patients, the BAF cytogram was distinguished by the higher macrophage content and the increased number of NBT-positive cells in this population with the reduced neutrophil content in both blood and BAF. The deceased had elevated levels of IgM and IgA in the blood serum and BAF. The results of the study of cytokine production in MNC cultures in vitro indicate that the ratio of IFN-γ/IL-4 in the conditions of spontaneous cytokine production was higher in the deceased compared with the indicators of the surviving patients. This fact indicates the polarization of the immune response of the deceased in the direction of the cell type mediated by Th1 cells, which is confirmed by the peculiarities of the BAF cytokine profile in the deceased – the significant increase in the value of IFN-γ/IL-4 relative to the indicators in the survivors. Conclusion. The unfavorable outcome of COPD is associated with the increase in the number of neutrophil cells in BAF, in the blood, with the predominance of Th1-type activation of adaptive immunity at the local level and activation of both cellular and humoral mechanisms of adaptive immunity at the systemic level. The increased activity of innate immunity in COPD exacerbation, manifested by the increase in the number and metabolic activity of macrophages, is associated with the long-term survival of patients.

Connective Tissue Diseases on the Elderly Population in Dakar Hospital Setting

Introduction: Immunosenescence contributes to the development of auto-antibodies. However, while the prevalence of some autoantibodies increases with age, the incidence of connective tissue diseases decreases with age. This study aims to determine the clinical and paraclinical characteristics of connective tissue diseases in the elderly population. Materials and Methods: We conducted a retrospective and descriptive study, from March 2010 to March 2021, in the Internal Medicine Department of Aristide Le Dantec Hospital, including all the medical records of patients followed up for connective tissue disease and whose symptomatology began at an age greater than or equal to 65 years. Epidemiological, clinical, paraclinical, therapeutic and evolutionary data were collected from a pre-established survey form and then processed using SPSS software. Results: Overall, 22 cases were collected out of 275 seen on consultation. They involved 16 women and 6 men, aged between 65 and 85 years with a median of 70 years. The mean duration of the clinical picture was 15.7 months with extremes of 1 and 96 months. The clinical symptoms were joint pain in 21 patients associated with dry eye and mouth syndrome in seven cases and one patient underwent isolated dry eye and mouth syndrome. Joint involvement was deforming in 13 patients. Poor general condition was noted in 20 patients. Biology showed anemia in 14 patients, hyperleukocytosis (2 patients) and thrombocytosis (5 patients). The sedimentation rate (SR) was accelerated in 9 patients and the C-reactive protein (CRP) was positive in 12 patients. Immunology showed a positive Latex test (3 patients/6), Waaler-Rose reaction (8 patients/13), anti-cyclic citrullinated peptides (anti-CCP) antibodies (11 patients/11), anti-nuclear antibodies (2 patients/4). Anti-extractable nuclear antigens (anti-ENA) antibody testing in 4 patients showed positive anti-U1RNP (1 case), anti-SSA and anti-SSB (1 case). Plain X-Rays of the hands, wrists and feet showed destructive lesions in 16 patients. The diagnosis of rheumatoid arthritis (RA) was set in 21 patients, one of which was associated with Sjogren’s syndrome (SS) and one with primary Sjogren’s syndrome. Treatment was based on prednisone (21 cases), methotrexate (17 patients), and hydroxychloroquine (9 patients). The evolution was remarkable in 10 patients. Conclusion: Autoimmune diseases in the elderly are uncommon compared to the young adult population. In our study, we only found cases of rheumatoid arthritis and Sjogren’s syndrome. As the population ages, connective tissue diseases may be increasingly diagnosed.