Polymerase Chain Reaction For Diagnosis Research Articles

Molecular Study of (Mycoplasma Pneumoniae, Streptococcus Pneumoniae) in Respiratory Infections Patients in Thiqar Province

The high death and hospitalization rates caused by bacterial respiratory infections make them a major worldwide health problem. Acute otitis media, bacterial rhinosinusitis, pharyngotonsillitis, bronchitis, and community-acquired pneumonia are among the common respiratory tract illnesses caused by common bacterial respiratory infections, which include Streptococcus pneumoniae, Mycobacterium tuberculosis, Haemophilus influenzae, Chlamydia pneumoniae, Mycoplasma pneumoniae, and Legionella pneumophila. The study aimed to detect uncultivated bacteria in respiratory infections patients (Mycoplasma pneumoniae) and Streptococcus pneumoniae that caused respiratory infections and determined if it caused this infection. The study included 140 patients (67males and 73 females) who were suffering from respiratory infections, ages ranging from 10-80 years, in a period from 1 September 2023 to 1 February 2024, all of the 140 samples collected from Nasiriyah Teaching Hospital, Center for Chest Diseases and Respiratory Diseases in Nasiriyah, Souq Al-Shuyoukh General Hospital, Alhussein Teaching Hospital, Private Clinics in Shatrah. Specimens included a sputum sample and oropharyngeal swab for direct detecting by polymerase chain reaction (pcr) Diagnosis is based on the presence of the (16s rRNA) gene to detect (Mycoplasma pneumoniae, Streptococcus pneumoniae) patients had with un-cultivated bacteria where 3\140(2.14%) of patients had infection with Mycoplasma pneumoniae and 6\140(4.28%) with Streptococcus pneumoniae. The present study was conducted that the most detected bacterial in female group 6 (75.0%), while in the male group 2 (25.0%), in addition, the most isolated bacteria in female group were both M. pneumoniae 3 (50.0%), and S. pneumoniae 3 (50%),whil in age groups in first group 3, while non-detected bacteria in both fourth and seventh age group 0, fever and headache are the most abundant symptoms patients, followed by myalgia (90.6%), chill (66%), cough (59%), and dyspnea (36.6%).

Evaluation of Isolation and Polymerase Chain Reaction in Diagnosis of Mycoplasma Gallisepticum in Broiler Chickens in Kirkuk Governorate, Iraq

Evaluation of Isolation and Polymerase Chain Reaction in Diagnosis of Mycoplasma Gallisepticum in Broiler Chickens in Kirkuk Governorate, Iraq

Comparative Study of Colorimetric In Situ Hybridization and Quantitative Real-Time Polymerase Chain Reaction for Diagnosis of Infection by Leishmania infantum in Dogs in Formalin-Fixed and Paraffin-Embedded Skin.

The zoonotic visceral leishmaniasis is caused by the protozoan Leishmania infantum and dogs are reservoirs for this parasite. For the diagnosis of Leishmania at the species level in dogs in formalin-fixed, paraffin-embedded skin (FFPES) samples, colorimetric in situ hybridization (CISH) and quantitative real-time polymerase chain reaction (qPCR) are options, but their sensitivities are not well established. Therefore, the aim of this study was to determine the sensitivity of these two techniques in FFPES for the diagnosis of the L. infantum infection in dogs using culture as the reference standard. The FFPES of 48 dogs with cutaneous infection by L. infantum confirmed by culture and by multilocus enzyme electrophoresis were examined by CISH and qPCR using specific probes for L. infantum. The sensitivities of qPCR, CISH and their combination were, respectively, 77.0%, 58.0% and 83.3%. The sensitivities of qPCR in dogs with and without clinical signs were, respectively, 74.2% and 82.4%. The sensitivities of CISH in dogs with and without clinical signs were, respectively, 61.3% and 52.9%. The CISH and qPCR showed satisfactory sensitivities for the diagnosis of L. infantum in the FFPES of dogs, even in dogs without clinical signs, and their combination increases the sensitivity for this diagnosis.

Integrated Three-Dimensional Microdevice with a Modified Surface for Enhanced DNA Separation from Biological Samples.

Functional interfaces and devices for rapid adsorption and immobilization of nucleic acids (NAs) are significant for relevant bioengineering applications. Herein, a microdevice with poly(acrylic acid) (PAA) photosensitive resin was integrated by three-dimensional (3D) printing, named DPAA for short. Precise microscale structures and abundant surface carboxyl functional groups were fabricated for fast and high-throughput deoxyribonucleic acid (DNA) separation. Surface modification was then done using polydopamine (PDA) and poly(ethylene glycol) (PEG) to obtain modified poly(acrylic acid) (PAA)-based devices DPDA-PAA and DPEG-PAA rich in amino and hydroxyl groups, respectively. The fabricated device DPAA possessed superior printing accuracy (40-50 μm). Functionalization of amino and hydroxyl was successful, and the modified devices DPDA-PAA and DPEG-PAA maintained a high thermal stability like DPAA. Surface potential analysis and molecular dynamics simulation indicated that the affinity for DNA was in the order of DPDA-PAA > DPEG-PAA > DPAA. Further DNA separation experiments confirmed the high throughput and high selectivity of DNA separation performance, consistent with the predicted affinity results. DPDA-PAA showed relatively the highest DNA extraction yield, while DPEG-PAA was the worst. An acidic binding system is more favorable for DNA separation and recovery. DPDA-PAA showed significantly better DNA extraction performance than DPAA in a weakly acidic environment (pH 5.0-7.0), and the average DNA yield of the first elution was 2.16 times that of DPAA. This work validates the possibility of modification on integrated 3D microdevices to improve their DNA separation efficiency effectively. It also provides a new direction for the rational design and functionalization of bioengineering separators based on nonmagnetic methods. It may pave a new path for the highly efficient polymerase chain reaction diagnosis.

A-297 Usefulness of Non-Skin Samples in the PCR Diagnosis of Monkeypox

Abstract Background During 2022, human mpox cases were reported in several European countries. Until November 2022, in Spain, 7404 cases were detected, most of them linked to sexual intercourse. In that context, fast identification of cases and sources of infection were essential for an optimal management. Polymerase chain reaction (PCR) was the preferred laboratory test. Our objective was to evaluate the usefulness of sera and nasopharyngeal samples for MPXV detection, and its application in contact studies without skin or mucosal injuries. Methods In the Hospital Clínico Universitario of Zaragoza (Spain), 106 samples from 50 patients, 38 male and 12 female, aged between 11 to 62 years, with skin lesions suspected MPXV infection were analysed. Samples were collected between May and September 2022 in inactivating universal viral transport medium (Biocomma). Samples were obtained between 1 and 19 days of evolution. To perform the real-time PCR, the VIASURE Monkeypox Virus Real Time PCR Detection Kit and the VIASURE 48—Real Time PCR System were used. Nucleic acid extraction was performed with the automated nucleic acid extraction system MagLEAD12gC using the MagDEA®Dx SV reagents. Results Table 1 shows the results expressed in Ct values. Skin, anogenital and serum samples were the most efficient for MPXV diagnosis. Nevertheless, these data are influenced by the sample collection date. Conclusion Real-time PCR was positive in samples from all locations in 71,4% of the infected patients. Respiratory samples (pharyngeal and nasopharyngeal) were the ones that showed the highest number of false negative results, whereas there was only one skin sample and one serum sample with a negative result. This study shows that sera samples can be used in the diagnosis of MPXV infection because of ease of collection, applicability to contacts without lesions and the option of serology testing for other STI agents (HBV, HCV, HIV, Treponema pallidum…).

Incidence of herpes simplex virus type 2 positivity among women living with human immunodeficiency virus in South Africa.

For women living with HIV (WLHIV), co-infection with herpes simplex virus type 2 (HSV-2) causes severe genital ulcers and presents additional challenges for their HIV care. To inform preventive strategies, we aimed to determine the incidence and risk factors of HSV-2 positivity in a prospective cohort of South African women. The CAPRISA 002 study enrolled women at acute HIV infection between 2004 and 2020. HSV-2 testing was conducted by multiplex polymerase chain reaction (PCR) assay on collected vaginal swabs up to twice annually during follow-up. We calculated incidence as the number of new cases per 100 person-years (PYs) and used Cox-proportional-hazard regression to identify factors associated with time-to-HSV-2 PCR positivity. At enrolment, the median age of 171 women was 24years, interquartile range (IQR 21-28), and the estimated median days since HIV infection was 42 (IQR 22-65). Of participants tested at enrolment, HSV-2 antibody prevalence was 81.4% (105/129), and 10.6% (12/113) were positive by PCR. Among 147 women with a prior negative HSV-2 PCR diagnosis, we observed 47 new HSV-2 PCR positive cases over 424.4 PYs of follow-up, yielding an incidence rate of 11.1 cases per-100-PYs. HSV-2 PCR positivity incidence was higher among younger women (<25years: adjusted Hazard Ratio [aHR] = 5.91, 95%CI 3.02-11.6), those with bacterial vaginosis (BV) (Nugent score 7-10: aHR = 2.17, 95%CI 1.15-4.10) and lower CD4 counts (<500cells/μl: aHR = 2.04, 95%CI 1.08-3.87). After acute HIV infection in women, the incidence of HSV-2 PCR positivity was associated with younger age, BV diagnosis and lower CD4 count.

Plasmodium vivax Malaria in Duffy-Positive Patients in Rwanda.

Plasmodium vivax is the second-most common malaria pathogen globally, but is considered very rare in the predominantly Duffy-negative sub-Saharan African population. In 259 malaria patients from highland southern Rwanda, we assessed Plasmodium species and Duffy blood group status by polymerase chain reaction (PCR). Plasmodium falciparum, P. vivax, Plasmodium malariae, and Plasmodium ovale were seen in 90.7%, 8.1%, 11.6%, and 5.0%, respectively. Plasmodium vivax occurred more frequently as a monoinfection than in combination with P. falciparum. All P. vivax-infected individuals showed heterozygous Duffy positivity, whereas this was the case for only 3.1% of patients with P. falciparum monoinfection and malaria-negative control subjects (P < 0.01). Based on PCR diagnosis, P. vivax is not rare in southern Rwanda. All episodes of P. vivax were observed in heterozygous Duffy-positive patients, whereas elsewhere in Africa, P. vivax is also reported in Duffy-negative individuals. Refined mapping of Plasmodium species is required to establish control and elimination strategies including all malaria species.

Diagnostic efficacy of chest CT imaging in diagnosis of COVID-19 cases based on duration of symptoms.

Chest computed tomography (CT) imaging provides results more rapidly and with higher sensitivity than reverse transcription polymerase chain reaction in diagnosis of COVID-19. To evaluate diagnostic efficacy of chest CT imaging in diagnosis of COVID-19 cases based on age and duration of symptoms. A retrospective study conducted during December 2020 to June 2021 in a tertiary care hospital, India. Total 495 patients with typical clinical symptoms of COVID-19, reverse transcription polymerase chain reaction positive for COVID-19 and had undergone chest CT imaging were included. Descriptive statistical analysis was performed for all the variables. Receiver operating characteristic curve analysis was used to determine threshold value of chest CT severity score (CT_SS) based on duration of symptoms and age to diagnose COVID-19. Mean age of patients was 61.86 ± 10.77 years and 367 (71.4%) patients were male. Ground glass opacities were observed in 456 (92.1%) patients and in 332 (67.1%) patients, multilobes were affected. Total CT_SS showed positive correlation with age (r = 0.257) and duration of symptoms (r = 0.625). Total CT_SS >6 after a duration of 2 days of symptoms identified COVID-19 cases with sensitivity 90.8% (95% confidence interval [CI]: 87.5%-93.5%) and specificity 84.6% (95% CI: 76.2%-90.9%). Total CT_SS >11 in patients aged more than 60 years identified COVID-19 cases with sensitivity 47.4% (95% CI: 41.2%-53.6%) and specificity 87.3% (95% CI: 82.3%-91.4%). Threshold value of CT_SS determined will help to expedite diagnosis of COVID-19 patients by the clinicians in an early stage especially in India and other developing countries which have a high patient volume and limited health resources.

Evaluation of the Weeds around Capsicum annuum (CA) Cultivation Fields as Potential Habitats of CA-Infecting Viruses.

Capsicum annuum (CA) is grown outdoors across fields in Jeollabuk-do, South Korea. The weeds surrounding these fields were investigated regarding the infection of 11 viruses infecting CA during the year 2014-2018. In the reverse transcription polymerase chain reaction diagnosis, 546 out of 821 CA samples (66.5%) were infected by nine viruses, and 190 out of 918 weed samples (20.7%) were infected by eight viruses. Correlation analysis of the mutual influence of the viruses infecting CA and weeds during these 5 years showed that five viruses had significant positive correlations with the infection in both CA and weeds. Over the study period, the weeds infected by cucumber mosaic virus (CMV) in the previous year were positively correlated with the incidence of CMV infection in CA in the current year, although the correlation was lower for tomato spotted wilt virus (TSWV) compared to CMV. The CMV infection percent was 14.0% in summer annuals, 11.4% in perennials, and 7.8% in winter annuals. However, considering the overwintering period without CA, the infection percent was 5.2% higher in winter annuals and perennials than that in summer annuals, indicating that winter annual and perennial weeds served as the main habitats for insect vectors. The TSWV infection percent in weeds was 10.4% in summer annuals, 6.4% in winter annuals, and 6.2% in perennials. The weeds surrounding CA fields, acting as the intermediate hosts, were found to be the potent sources of infection, influencing the spread and diversity of CA-infecting viruses. The results of this study can contribute to prevent viral infection in agricultural fields.

Impact of malaria diagnostic choice on monitoring of Plasmodium falciparum prevalence estimates in the Democratic Republic of the Congo and relevance to control programs in high-burden countries.

Malaria programs rely upon a variety of diagnostic assays, including rapid diagnostic tests (RDTs), microscopy, polymerase chain reaction (PCR), and bead-based immunoassays (BBA), to monitor malaria prevalence and support control and elimination efforts. Data comparing these assays are limited, especially from high-burden countries like the Democratic Republic of the Congo (DRC). Using cross-sectional and routine data, we compared diagnostic performance and Plasmodium falciparum prevalence estimates across health areas of varying transmission intensity to illustrate the relevance of assay performance to malaria control programs. Data and samples were collected between March-June 2018 during a cross-sectional household survey across three health areas with low, moderate, and high transmission intensities within Kinshasa Province, DRC. Samples from 1,431 participants were evaluated using RDT, microscopy, PCR, and BBA. P. falciparum parasite prevalence varied between diagnostic methods across all health areas, with the highest prevalence estimates observed in Bu (57.4-72.4% across assays), followed by Kimpoko (32.6-53.2%), and Voix du Peuple (3.1-8.4%). Using latent class analysis to compare these diagnostic methods against an "alloyed gold standard," the most sensitive diagnostic method was BBA in Bu (high prevalence) and Voix du Peuple (low prevalence), while PCR diagnosis was most sensitive in Kimpoko (moderate prevalence). RDTs were consistently the most specific diagnostic method in all health areas. Among 9.0 million people residing in Kinshasa Province in 2018, the estimated P. falciparum prevalence by microscopy, PCR, and BBA were nearly double that of RDT. Comparison of malaria RDT, microscopy, PCR, and BBA results confirmed differences in sensitivity and specificity that varied by endemicity, with PCR and BBA performing best for detecting any P. falciparum infection. Prevalence estimates varied widely depending on assay type for parasite detection. Inherent differences in assay performance should be carefully considered when using community survey and surveillance data to guide policy decisions.

Finger printing-RFLP analysis of chromosomal IS6110 insertion sequence and PCR diagnosis of pulmonary tuberculosis, isolated from patients in El Hajeb region of Morocco

Tuberculosis (TB) is one of the contagious diseases caused by M. tuberculosis (MTB) bacteria. Prompt diagnosis is one of the active solutions to control the spread of this infection. Besides, a targeted, specific and non-complex diagnosis can prove promising in this type of epidemic. This study was designed to compare the efficiencies of a diagnosis by Ziehl-Neelsen staining (ZN) and by the polymerase chain reaction (PCR) technique. Samples presented smear-positive pulmonary TB were subjected to Chromosomal restriction fragment length polymorphism of IS6110 (IS6110-RFLP) for fingerprinting profile determination.The results showed that out of 100 sputum samples of suspected case, 53 were positive. Numbers of positive individuals for tuberculosis obtained by the different diagnostic techniques, to know, (ZN staining; culture and PCR) were respectively: 6, 25 and 22. Chromosomal RFLP fingerprinting profile revealed the presence of five different genotypes obtained from seven tested isolates.These results suggest that molecular techniques are alternative tool for fast and specific diagnosis of pulmonary MTB from sputum.

Diagnostic of visceral leishmaniasis by polymerase chain reaction PCR in experimental mice infected which treatment with Viscum album extract and Pentostam.

Visceral leishmaniasis (VL), or kala-azar, is an infection of the reticuloendothelial system, Fatalities due to leishmaniasis are associated with visceral disease. the Leishmaia donovani is a causative agent for this disease. In the current study, white male mice of the Balb /C strain were experimentally infected with the leishmania donovani promastigotes and were then treated with a Viscum extract and compared with the Pentostam. The aim of this study was to evaluate the sensitivity and specificity of polymerase chain reaction (PCR) in the detection of Leishmania DNA in blood samples of infected mice. as well as, evaluation the ability of Viscum album extract to treatment the visceral leishmaniasis. The genetic study gave excellent results in the diagnosis of infection throughout the treatment period compared to the positive and negative control . Using the specific LdI primer , the 700bp of the parasite was diagnosed. This Broad band was shown in the positive control samples, in the pantostam treatment groups, and the muscular injection with viscum extract in the first 15 days and in the oral dosage group in the first 20 days of initiation. While the band did not appear in any other. The current study has shown promising results in the use of V. album in the treatment of visceral leishmaniasis and also gave impressive results in the diagnosis of infection PCR.

Strong Association between Vitamin D Receptor Gene and Severe Acute Respiratory Syndrome coronavirus 2 Infectious Variants.

A coronavirus disease 2019 (COVID-19) disease, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has created significant concern since December 2019 worldwide. The virus is known to be highly transmissible. Heterogenic clinical features even vary more among SARS-CoV-2 variants from asymptomatic forms to severe symptoms. Previous studies revealed an association between COVID-19 and vitamin D deficiency resulting from its low levels in COVID-19 patients. To our knowledge, there is no scientific investigation that evaluates the direct association between SARS-CoV-2 variants of concern and vitamin D receptor ( VDR ) gene markers in Cyprus. Thus, the present study aimed to identify the putative impact of VDR gene polymorphisms on SARS-CoV-2 infection among different variants. The nasopharyngeal swabs were taken from a total number of 600 patients who were admitted to Near East University Hospital COVID-19 Polymerase Chain Reaction (PCR) Diagnosis Laboratory for routine SARS-CoV-2 real-time quantitative reverse transcription PCR (RT-qPCR) test. The RT-qPCR negative resulting samples were taken as control samples ( n = 300). On the contrary, the case group consisted of patients who were SARS-CoV-2 RT-qPCR positive, infected with either SARS-CoV-2 Alpha ( n = 100), Delta ( n = 100), or Omicron ( n = 100) variants. Two VDR gene polymorphisms, Taq I-rs731236 T > C and Fok I-rs10735810 C > T, were genotyped by polymerase chain reaction-restriction fragment length polymorphism. The mean age of the COVID-19 patient's ± standard deviation was 46.12 ± 12.36 and 45.25 ± 12.71 years old for the control group ( p > 0.05). The gender distribution of the patient group was 48.3% female and 51.7% male and for the control group 43% female and 57% male ( p > 0.05). Significant differences were observed in genotype frequencies of FokI and TaqI variants between SARS-CoV-2 patients compared to the control group ( p < 0.005). Furthermore, the risk alleles, FokI T allele and TaqI C, were found to be statistically significant (odds ratio [OR] = 1.80, 95% confidence interval [CI] = 1.42-2.29, OR = 1.62, 95% CI = 1.27-2.05, respectively) in COVID-19 patients. The highest number of patients with wild-type genotype was found in the control group, which is 52.9% compared with 17.5% in the case group. Moreover, most of the COVID-19 patients had heterozygous/homozygous genotypes, reaching 82.5%, while 47.1% of the control group patients had heterozygous/homozygous genotypes. Our results suggested that patients with FokI and TaqI polymorphisms might tend to be more susceptible to getting infected with SARS-CoV-2. Overall, findings from this study provided evidence regarding vitamin D supplements recommendation in individuals with vitamin D deficiency/insufficiency in the peri- or post-COVID-19 pandemic.

Comparative assessment of regulated methods and PCR in the diagnosis of trichophytosis in veterinary mycology.

There is an increase in the incidence of human and animal infectious skin diseases of fungal etiology in the world. The main source of infecting the population has become agricultural and stray animals. The objective of this study was to examine the morphophysiological and microbiological characteristics of pathogenic fungi belonging to the species Trichophyton verrucosum. This species is known to cause diseases in both humans and livestock in Kazakhstan. In addition, the study aimed to assess the feasibility of using the polymerase chain reaction (PCR) method for detecting T. verrucosum. This assessment was conducted in comparison to the outcomes of conventional laboratory diagnostic tests commonly employed for trichophytosis. The research focused on analyzing 141 samples of pathological material obtained from calves in Almaty, Turkestan, and Kyzylorda regions. These calves exhibited clinical symptoms of skin disease. The study aimed to identify the causative agent using various techniques, including microscopic examination, microbiological methods involving the isolation of pure cultures, and PCR. The detection of the causative agent of dermatophytosis using conventional methods was relatively low, 86% for the microscopic method, and 79% for the microbiological method with the isolation of the culture of the pathogen. Extraction and detection of the genetic material of the causative agent of the disease for PCR was carried out according to the method developed by the authors. The effectiveness of the PCR method was 97.9%, which is significantly higher (p < 0.05) compared with the diagnostic effectiveness of conventional methods. The PCR method using specific primers identified the causative agent in 98% of cases, which significantly (p < 0.05) exceeded the results obtained using conventional diagnostic methods. Accordingly, the PCR method had better sensitivity and specificity indicators. The conducted study recommends the method of PCR diagnosis of dermatophytosis for fast and reliable confirmation of the diagnosis of dermatophytosis in humans and animals in Kazakhstan.

Sensitivity of PCR in conjunctival swab samples for the diagnosis of canine visceral leishmaniasis: a systematic review and meta-analysis.

To compare the sensitivity of conjunctival swab (CS) and conventional samples (blood, spleen, liver, lymphoid and cutaneous tissue) in the diagnosis of canine visceral leishmaniasis (CVL) by polymerase chain reaction (PCR), a systematic review and meta-analysis was carried out using PubMed, Science Direct, Scopus, Web of Science, VHL/BVS (Virtual Health Library), CAPES, and Scielo databases. Articles published from 2002 to 2022 were considered and the review was updated in Jul 2023. From the total of 371 identified studies, 8 met all the eligibility criteria and were included in this review. Data from 658 CVL-positive dogs and 2541 PCR results were considered. Using a random effect model, data on the sensitivity of the test was compared between intervention (CS samples) and comparison (all the other samples) groups. Overall, the use of CS in the PCR diagnosis of CVL produced 12% higher sensitivity (p=0.013) in the test than all the other samples in combination. The animals' clinical condition did not influence (p>0.142) this overall result. However, when CS was individually compared to each of the conventional samples, the consistent result was observed (p=0.012) only in the CS versus bone marrow comparison. Given their rapid acquisition, minimal invasiveness, and lower cost relative to conventional samples, CS samples present a promising alternative for the molecular diagnosis of CVL.

Accuracy of Diagnostic Tests for the Detection of Chagas Disease: A Systematic Review and Meta-Analysis.

The present systematic review and meta-analysis about the accuracy of diagnostic tests aim to describe the findings of literature over the last thirty years for the diagnosis of Chagas disease (CD). This work aimed to determine the accuracy of diagnostic techniques for CD in the disease's acute and chronic phases. The PubMed database was searched for studies published between 1990 and 2021 on CD diagnostics. Fifty-six published studies that met the criteria were analyzed and included in the meta-analysis, evaluating diagnostic accuracy through sensitivity and specificity. For Enzyme-Linked Immunosorbent Assay (ELISA), Fluorescent Antibody Technique (IFAT), Hemagglutination Test (HmT), Polymerase Chain Reaction (PCR), and Real-Time Polymerase Chain Reaction (qPCR) diagnosis methods, the sensitivity had a median of 99.0%, 78.0%, 75.0%, 76.0%, and 94.0%, respectively; while specificity presented a median of 99.0%, 99.0%, 99.0%, 98.0%, and 98.0%, respectively. This meta-analysis showed that ELISA and qPCR techniques had a higher performance compared to other methods of diagnosing CD in the chronic and acute phases, respectively. It was concluded utilizing the Area Under the Curve restricted to the false positive rates (AUCFPR), that the ELISA diagnostic test presents the highest performance in diagnosing acute and chronic CD, compared to serological and molecular tests. Future studies focusing on new CD diagnostics approaches should be targeted.

Monitoring Leishmania infantum Infections in Female Lutzomyia longipalpis by Using DNA Extraction on Cation Exchange Paper and PCR Pool Testing

Visceral leishmaniasis remains a serious public health issue, and Brazil was among the seven countries with the highest prevalence of this disease worldwide. The measures to control this disease are not easily developed, and the improvement of its diagnosis, surveillance, and control is still needed. This study aimed to carry out the polymerase chain reaction (PCR) diagnosis of Leishmania infantum in vector samples in some municipalities of the State of São Paulo, which included two municipalities with human disease transmission and two with dog transmission only. Vectors were collected in traps with luminous bait. Next, they were killed at -4 °C and kept in 70% alcohol. Groups of ten female insects (pools) were mashed on cation exchange paper (fine cellulose phosphate with 18 µEq/cm² ionic exchange capacity) for DNA extraction. The PCR was carried out to identify the natural infection of the Leishmania genus in female Lutzomyia longipalpis (Lu. Longipalpis). Out of the 3,880 Lu. longipalpis phlebotomines, 1060 were female and 2820 were male (3:1). The method used to extract the DNA in pools of ten phlebotomines and the PCR resulted in sensitivity, specificity, practicality, and faster analyses when compared to the individual analysis method. The procedure described can be used on a large scale in the leishmaniasis epidemiological surveillance, enabling a higher number of analyses and the optimization of human resources because the traditional diagnostic method is carried out via desiccation of the insect digestive system and microscopic examination, which is time-demanding and there is the need of manual skills.

Application of Multiplex Real-Time Polymerase Chain Reaction in Diagnosis of Intestinal Parasitic Infection among Anemic Pregnant Women

Background: Intestinal parasitic infection can cause anemia in pregnant women and is a major public health problem. Ascariasis, amebiasis, hookworm, giardiasis, and trichuriasis are common intestinal infestations causing anemia. The study detects them by using multiplex real-time polymerase chain reaction (PCR). Methods: Stool samples were collected, transported, DNA extraction was done followed by multiplex real-time PCR was performed using specific primers and probes for their detection. The results so obtained were compared with results of microscopy. Results: A total of 234 pregnant women were enrolled in the study, of which 82.5% were anemic. Among them, parasites were detected by multiplex real-time PCR such as Ascaris lumbricoides in 73%, Ancylostoma duodenale in 8%, Entamoeba histolytica in 17%, Giardia duodenalis in 2%, and Trichuris trichiura 0%. All pregnant women with intestinal parasitic infection were anemic. Multiplex real-time PCR showed a high detection rate as compared to microscopy (100% vs. 14.7%). Polyparasitism was detected in 11% of cases using multiplex real-time PCR. Conclusion: Multiplex real-time PCR enabled the detection of intestinal parasites in pregnant anemic women with distinct advantage over microscopy. It can potentially be used in an epidemiological surveillance study and can potentially influence the treatment in practice for intestinal parasitic infection causing anemia in pregnant women.

Biomarkers predicting the 30‑day mortality of patients who underwent elective surgery and were infected with SARS‑CoV‑2 during the post‑operative period: A retrospective study.

The coronavirus disease 2019 (COVID-19) pandemic is a significant global concern that has had major implications for the healthcare system. Patients with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) undergoing elective or emergency surgical procedures have a substantial risk of mortality and peri-operative complications. The present study aimed to describe the characteristics of patients who underwent elective surgery and developed nosocomial SARS-CoV-2 infection post-surgery. Patients who underwent thoracic, upper and lower abdominal or peripheral elective surgery with a polymerase chain reaction diagnosis of COVID-19, at 3-7 days after the surgery, were enrolled in the present retrospective study. Demographics, vaccination status against SARS-CoV-2, Charlson comorbidity index (CCI) and laboratory data were recorded upon admission to the hospital unit. In total, 116 subjects (80 males, 36 females; mean age, 67.31±16.83 years) fulfilling the inclusion criteria were identified. Among the 116 participants, 14 (12.1%) were intubated. From the 116 individuals analyzed, 84 were alive after 30 days (survivors), and 32 had succumbed to the disease (non-survivors). The mortality rate was 27.6% (32/116). The non-survivors had an older age and a higher CCI score. At the evaluation upon admission to the hospital unit, the survivors presented with higher serum albumin levels and a higher number of blood lymphocytes. In addition, the survivors exhibited lower levels of lactate dehydrogenase, aspartate aminotransferase, alkaline phosphatase (ALP) and C-reactive protein (CRP), as well as a higher neutrophil to lymphocyte ratio (NLR) and CRP to albumin ratio (CAR) (P<0.05). The patients that were intubated had higher levels of gamma glutamyl-transferase (GGT), ALP and ferritin, as well as a higher NLR and platelet to lymphocyte ratio upon admission to the hospital unit (P<0.05). According to the Cox proportional hazards multivariate regression analysis, the only independent predictors of mortality and intubation were ALP and GGT upon admission, respectively (P<0.05). On the whole, the findings of the present study suggest that more stringent guidelines are required in order to prevent infection during the post-operative period.

Characteristics of polymerase chain reaction-positive syphilis cases in Manitoba, Canada, 2017 to 2020: Demographic analysis, specimen types, and Treponema pallidum gene targets.

A resurgence of syphilis infections has been described in a number of countries including Canada in the last decade. This study identified polymerase chain reaction (PCR) positive syphilis cases based on detection of Treponema pallidum genes (polA, tpp47, and bmp) in 3,350 clinical specimens obtained from patients in the province of Manitoba, Canada between 2017 and 2020. Patient demographics were obtained from specimen requisition forms. PCR identified 740 syphilis cases: 718 were adolescents and adults, while 22 were congenital syphilis cases. For non-congenital syphilis investigation, the clinical specimens with the highest yield of positive PCR results were genital (632), oral (73), and anal (55), while for congenital syphilis, they were nasal or nasopharyngeal secretions (20), followed by blood (5) and umbilical cord (4). Female syphilis cases appeared younger (61.7% between 14 and 29 years), while male syphilis cases appeared older (58.4% between 30 and 65 years). Although, overall more syphilis cases (62.7%) occurred in the urban cities; the proportion of urban cases showed a significant decline from 87.0% in 2017 to 55.6% in 2020, while in rural regions it increased from 13.0% in 2017 to 44.4% in 2020. Most (98.8%) PCR- positive specimens were found to contain all three T. pallidum genes and 99.8% also displayed the macrolide resistance genotype. This study identified the clinical specimen types and T. pallidum genes most suitable for PCR diagnosis of syphilis. Changing demographics of cases were noted over time.