Rapid Immunochromatographic Test Research Articles

Hepatitis B virus infection among illegal drug users in Enugu State, Nigeria: prevalence, immune status, and related risk factors

IntroductionHepatitis B virus infection poses a global health challenge, particularly in low- and middle-income African countries. Illicit drug use exacerbates the problem, with drug users having a higher HBV infection risk and maintaining a quiet transmission pool. This study aimed to determine HBV infection prevalence, immune status, and risk factors among illegal drug users in Enugu State, Nigeria.Materials and methodsA cross-sectional study was conducted in Enugu State, using privileged access interviewer methods to enroll drug users. Pre-tested structured questionnaires were administered after informed consent was obtained. Blood samples were tested for HBsAg, HBsAb, HBeAg, HBeAb, and HBcAb using immunochromatographic rapid test kits. Statistical analysis was performed using SPSS version 25.ResultsOne hundred drug users were recruited into the study. Overall, 7% of the drug users had HBV infection. 61% were found to be susceptible to HBV infection, 15% showed serological evidence of immunity from HBV vaccination and 1% developed natural immunity from HBV exposure. Significant risk factors for HBV infection were age less than 20 years, young age (≤ 20 years) at drug initiation, being single, injecting drugs more than or equal to 20 times per month and injecting with used syringes.ConclusionThis study shows high intermediate endemicity of HBV infection among drug users, low vaccination uptake, and high susceptibility to HBV infection. This calls for the urgent inclusion of drug users in national HBV vaccination campaigns and the adoption of the World Health Organization’s recommendations on the prevention of viral hepatitis among people who inject drugs.

Study of association of thrombocytopenia and serological parameters in dengue fever with special reference to NS1 antigen in hilly Region of Northern India

Background: Dengue fever is an arthropod-borne disease transmitted to humans by Aedes aegypti mosquitoes and one of the leading causes of arthropod-borne viral disease in the world. Aims and Objectives: The present study aims to analyze any association between platelet count and immunochromatography -based dengue serology tests. Materials and Methods: This was a cross-sectional study conducted in Government Doon Medical College and Hospital. A total of 19,208 clinically suspected cases of dengue who reported in various outpatient departments, emergency services, and inpatient departments of our hospital between June 2023 and November 2023 were included in this study. Samples were tested by rapid immunochromatographic test for dengue non-structural protein 1 (NS1) antigen, immunoglobulin M (IgM), IgG, and platelet count was also obtained and a comparison was made. Results: Out of total 19,208 samples tested, 4876 (25.38%) samples were positive for NS1, 589 (3.06%) samples were positive for IgM. Majority of the patients were in the age group of 21–40 years 9274 (48.28%). 11,753 (61.19%) were males and 7455 (38.80%) were females. A strong correlation was seen between NS1 and thrombocytopenia where a total of 2737/4876 (n=56.13%) patients showed positivity in NS1 with low platelet counts. Conclusion: Detection of NS1 antigen helps in early diagnosis of dengue to avoid complications significantly. In confirmed dengue cases with fever, thrombocytopenia is more consistently found and can be used as a predictor to reduce the morbidity and mortality of dengue disease.

Diagnostic performance of a rapid immunochromatographic test for the simultaneous detection of antibodies to Theileria equi and Babesia caballi in horses and donkeys

BackgroundEquine piroplasmosis is caused by two tick-borne protozoan parasites, Theileria equi and Babesia caballi,, which are clinically relevant in susceptible horses, donkeys, and mules. Moreover, equine piroplasmosis significantly constrains international trading and equestrian events. Rapidly diagnosing both parasites in carrier animals is essential for implementing effective control measures. Here, a rapid immunochromatographic test for the simultaneous detection of antibodies to T. equi and B. caballi was evaluated using samples from horses and donkeys collected in Greece, Israel, and Italy. The results were compared with an improved competitive enzyme-linked immunosorbent assay (cELISA) for detecting antibodies to both parasites using the same panel of samples.MethodsBlood samples were collected from 255 horses and donkeys. The panel consisted of 129 horses sampled at four locations in northern Greece, 105 donkeys sampled at four locations in Sicily, and 21 horses sampled at two locations in Israel. The rapid test and the cELISA were performed according to the manufacturer’s instructions, and the results were subjected to a statistical analysis to determine the sensitivity and specificity of both tests and their association.ResultsThe immunochromatographic test provided a result within 15 min and can be performed in the field, detecting both pathogens simultaneously. The overall coincidence rate between the rapid test and the cELISA for detecting antibodies against T. equi was 93% and 92.9% for B. caballi. The rapid test’s sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for T. equi were above 91.5%. Sixteen samples were positive for both parasites in the rapid test and eight in the cELISA. Either test had no significant association between T. equi and B. caballi detection. The detection rates of both parasites were significantly higher in Italy than in Greece or Israel and in donkeys than in horses. The agreement for T. equi between the results of both tests was high in Greece (93.8%) and Italy (95.2%) and moderate in Israel (76.2%). For B. caballi, the specificity and NPV of the rapid test were high (94.2% and 98.3%, respectively), although the sensitivity and PPV were moderate (69.2% and 39.1%, respectively) due to the small sample size. However, for B. caballi, the sensitivity was higher with the rapid test.ConclusionsThe rapid test detected T. equi and B. caballi simultaneously in the field, potentially replacing laborious cELISA testing and is recommended for import/export purposes. The test can also be helpful for the differential diagnosis of clinical cases, since seropositivity may rule out equine piroplasmosis since it does not indicate current or active infection.Graphical

QUANTIFICATION OF HEPATITIS B AND HEPATITIS C VIRAL LOAD BY REAL-TIME PCR AND ASSESSMENT OF COINFECTION OF HBV AND HCV AMONG HIV-POSITIVE PATIENTS IN A TERTIARY CARE HOSPITAL

bjective: To assess the prevalence of coinfection of HIV-HBV and HIV-HCV and to estimate the viral load of HBV and HCV in PLWH.
 Methods: A retrospective study was conducted from November 2021 to October 2022 among patients attending ICTC. 5 ml of blood sample collected aseptically was tested for HIV, HBV and HCV using rapid immunochromatographic tests, ELISA and viral load estimated by Real-time PCR.
 Results: Out of 5087 samples tested for HIV, 666 samples (13.09%) were found to be positive. Prevalence of HIV-HBV and HIV-HCV coinfection was 15.6%(104 cases) and 1.5% (10 cases), respectively. Out of which, males were predominant (62.28%). This is clinically significant with a p-value of * 0.05. HIV-HBV and HIV-HCV coinfections were predominant in 41-50 y age group. Among104 HIV-HBV coinfected, viral load at the time of diagnosis is ‘below detection level’ in 25(24.04%), <250 in 15(14.42%), 251-500 in 9(8.65 %), 501-1000 in 13(12.5 %), 1001-10,000 in 23(22 %) and>10, 000 copies/ml in 19(18.26%). In 10 HIV-HCV coinfected cases, the viral load is ‘below detection level’ in 2(20%), <250 in 1(10%), 251-500 in 1(10 %), 501-1000 in 1(10 %), 1001-10,000 in 4(40%) and>10,001-100,000 copies/ml in 1(10%).
 Conclusion: Monitoring the viral load in HBV or HCV infected at the time of diagnosis of HIV or testing for protective levels of antibodies post-vaccination in uninfected people will help in limiting the progression of chronic HBV or HCV to cirrhosis, end-stage liver disease or hepatocellular carcinoma.

Refinement of the rKLi8.3-Based Serodiagnostic ELISA Allows Detection of Canine Leishmaniosis in Dogs with Low Antibody Titers.

The diagnosis of canine leishmaniasis (CanL) still represents a challenge due to the variable clinical manifestations and the large number of asymptomatic dogs. Serological tests are most commonly used to detect infected animals, revealing anti-Leishmania antibodies, mainly of the IgG isotype. Recently, a new diagnostic antigen, rKLi8.3, containing 8.3 kinesin tandem repeats (TR) from a Leishmania infantum strain from Sudan, has been shown to provide excellent specificity and sensitivity for the detection of Leishmania-infected humans and dogs. However, asymptomatic animals with very low antibody titers are often difficult to detect by serodiagnosis. Thus, we wondered whether the addition of an anti-IgG-enhancing step in the protein A/G-based rKLi8.3-ELISA will improve the diagnostic performance without decreasing the specificity. For this, parasitologically confirmed CanL cases with low or high clinical scores, uninfected healthy controls and dogs with other infections were tested by rKLi8.3-ELISA as well as two different immunochromatographic rapid tests, rKLi8.3-lateral flow test (LFT) and Dual Path Platform (DPP®) based on the rK28 antigen. Our results show that the diagnostic accuracies of the rKLi8.3-ELISA and LFT were similar to that of DPP, missing several asymptomatic animals. However, the addition of a secondary, amplifying anti-dog IgG antibody in the protein A/G-based rKLi8.3-ELISA enabled the detection of nearly all asymptomatic dogs without compromising its specificity.

A rapid and sensitive time-resolved fluorescence microsphere immunochromatographic test strip for bongkrekic acid detection in edible fungi

Bongkrekic acid (BA) is a prevalent biological toxin in black fungus, tremella and fermented rice flour food. BA usually causes acute poisoning. Developing a rapid on-site detection method for BA is essential for timely prevention. This work proposed a time-resolved fluorescent microsphere immunochromatographic test strip (TRFM-ICTS) for the quantitative determination of BA in edible fungi. The process involved synthesizing BA artificial antigens and fluorescent probes. An independent quality control system was implemented. This system ensured the integrity of the control line (C line) even when the test line (T line) was present. Under the optimal conditions, the linear range of TRFM-ICTS was 0.1–10 ng/mL, and the half inhibitory concentration (IC50) was 0.264 ng/mL. The detection limit of the developed test strip (0.282 ng/mL) was 20 times lower than that of colloidal gold immunochromatographic test strip (5.7 ng/mL). Moreover, TRFM-ICTS showed good recovery in spiked samples, ranging from 81.22% to 119.48%, and the coefficient of variation (CV) was less than 7.76%. In comparison to conventional instrumental techniques, the implemented TRFM-ICTS demonstrated a highly sensitive, straightforward, expeditious, and feasible approach for the quantification of BA in food.

Visceral leishmaniasis seroprevalence and associated sociocultural factors in an endemic area of Sergipe, northeastern Brazil

The present study aimed to investigate the presence of anti-Leishmania antibodies in samples of domiciled dogs and associate them with epidemiological indicators in the city of Lagarto, Sergipe. The diagnosis of Canine Visceral Leishmaniasis (CVL) was carried out in 755 dogs, where the rapid two-way platform immunochromatographic test (TR-DPP) (Fiocruz/Biomanguinhos) and the serological test of Enzyme-Linked Immunosorbent Assay (ELISA) were applied. The results showed that all homes with seroreactive dogs were close to abandoned land, squares and/or green areas. There was a significant association between seroactive animals and type of shelter (p=0.008); 54.8% had an indoor shelter and 45.2% of the dogs had a shelter outside their homes. The disease is geographically distributed in 14 (73.68%) of the 19 sample areas in the municipality. Of these, five (26.31%) had cases of Visceral Leishmaniasis in dogs and humans. Eight cases of Human Visceral Leishmaniasis (HVL) were reported during the years 2017 to 2020 in the municipality. Dogs that are located in areas of greatest risk impact the maintenance of natural foci of Leishmaniasis transmission to human and animal hosts in areas endemic for Visceral Leishmaniasis. Such results are essential to assist epidemiological surveillance in the municipality of Lagarto-SE with the implementation of control measures aimed at the insect vector of the disease, the sand fly, to prevent human and canine cases.

Rotavirus and adenovirus infections in children with acute gastroenteritis after introducing the Rotasiil® vaccine in Kisangani, Democratic Republic of the Congo.

Although rotavirus vaccination has reduced the global burden of the virus, morbidity and mortality from rotavirus infection remain high in Sub-Saharan Africa. This study aimed to determine the prevalence of rotavirus and adenovirus infections in children under five years with acute gastroenteritis and to identify factors associated with rotavirus infection after the introduction of the Rotasiil® vaccine in 2019 in Kisangani, Democratic Republic of the Congo (DRC). This study consisted of a cross-sectional hospital-based survey conducted from May 2022 to April 2023 in four health facilities in Kisangani, using a fecal-based test (rapid antigenic immuno-chromatographic diagnostic test, BYOSYNEX adenovirus/rotavirus BSS, Biosynex SA, Illkirch-Graffenstaden, France) of rotavirus and adenovirus infections among children under five years of age with acute gastroenteritis. A total of 320 children under five years of age with acute gastroenteritis were included. The prevalence of rotavirus infection was 34.4%, that of adenovirus was 6.3%, and that of both rotavirus and adenovirus coinfection was 1.3%. The prevalence of rotavirus was significantly higher in unvaccinated children than in vaccinated children (55.4% versus 23.1%; P < 0.001). This difference was observed only in children who received all three vaccine doses. Multivariate logistic regression analysis shows that the rate of rotavirus infection was significantly reduced in vaccinated children (adjusted OR: 0.31 [95% confidence intervals (CI): 0.19-0.56]; P < 0.001) and those whose mothers had an average (adjusted OR: 0.51 [95% CI: 0.25-0.91]; P = 0.018) or high level (adjusted OR: 0.34 [95% CI: 0.20-0.64]; P < 0.001) of knowledge about the rotavirus vaccine. The prevalence of rotavirus infection remains high in Kisangani despite vaccination. However, the prevalence of adenovirus infections was low in our series. Complete vaccination with three doses and mothers' average and high level of knowledge about the rotavirus vaccine significantly reduces the rate of rotavirus infection. It is, therefore, essential to strengthen the mothers' health education, continue with the Rotasiil® vaccine, and ensure epidemiological surveillance of rotavirus infection.

Risk factors for the presence of antibody against anaplasma spp. In dogs in Poznan.

The aim of the study was to identify the incidence of antibodies against Anaplasma spp. in dogs living in Poznań. Canine anaplasmosis is a tick-borne dog disease caused by Anaplasma phagocytophilum or A. platys infection. To identify the prevalence of antibodies against Anaplasma spp. in dogs, we analyzed blood samples collected from 349 client-owned dogs living in Poznań (Poland). Using an immunochromatographic rapid test (Caniv-4, VetExpert, Poland), antibodies against Anaplasma spp. were detected in 32 (9.2%) dogs aged over two years. Sex, hair length, and location (region of Poznań) were not identified as statistically significant risk factors for the presence of antibodies against Anaplasma spp. The odds ratio for antibodies against Anaplasma spp. in large breed dogs vs small breed dogs was 3.76. The results of the study suggest a growing presence of Anaplasma spp. in dogs living in big cities in Poland.

Nitazoxanide refractory cryptosporidiosis complicating Burkitt lymphoma in a child.

Cryptosporidium species cause watery diarrhea in several vertebrate hosts, including humans. Most apparently, immunocompetent-infected individuals remain asymptomatic, whereas immunocompromised may develop severe or chronic cryptosporidiosis. We report here the case of a 6-year-old girl undergoing chemotherapy for Burkitt lymphoma who experienced multiple episodes of watery diarrhea during her hospital stay. Microscopic examination of her stool sample revealed oocysts of Cryptosporidium species. The rapid immunochromatographic test was also positive for Cryptosporidium species. She was treated with nitazoxanide for 3 weeks, which failed to provide both clinical improvement and parasitological clearance. This case highlights the importance of treatment failure in human cryptosporidiosis.

Giant chancre in an octogenary patient. Always remember syphilis!

Introduction: Syphilis is a sexually transmitted infection (STI) caused by the Treponema pallidum and has become epidemic in the past two decades. The disease is often asymptomatic, and its manifestations are diverse. It may affect multiple systems, posing the risk of serious consequences or even death. Objective: To present a case of an exuberant syphilis-related lesion in an elderly man. Case report: An 85-year-old man presented with a painless penile lesion. He reported having had oral sexual contact (felatio) with a female sex worker six weeks prior the presentation of the lesion. During the physical exam, an ulcer measuring three to four cm was observed, reaching subcutaneous tissue down to the Buck’s fascia, and presenting a clean beefy red base, and an infiltrated edge. Bilateral and movable enlarged and painless lymph nodes were found. The rapid immunochromatographic test was positive for syphilis and negative for HIV, hepatitis B, and hepatitis C. Treatment consisted of an in loco injection of 2,400,000 IU of benzathine penicillin. Conclusion:In spite of being more common among younger individuals, syphilis should be always considered in the differential diagnosis of genital ulcers, regardless of the patient’s age. Many aged people keep an active sexual life especially with the increase of longevity and effective therapeutic measures for erectile disfunction. The false belief that sexual activity ceases to exist in older age groups must, therefore, be abandoned.

Human rickettsial infections in India - A review.

Rickettsial infections are emerging and/or re-emerging disease that poses a serious global threat to humans and animals. Transmission to humans and animals is through the bite of the ectoparasites including ticks, fleas and chigger mites. Most of the rickettsial diseases are endemic in India, but underdiagnosed. This review is aimed at analyzing the prevalence of rickettsiosis in India and the advancement of rickettsial diagnosis. We have conducted a systematic review on the prevalence of rickettsial disease in India ranging from 1.3% to 46.6% for spotted fever, 2.4% to 77.8% for scrub typhus and 1% to 46.4% for Q fever, based on the literature published with the evidence of isolation, serological, and molecular diagnostics. Search engines Medline/PubMed, Science Direct, ProQuest, and EBSCO were used to retrieve the articles from electronic databases by using appropriate keywords to track the emergence of these rickettsial diseases in India for the period of 1865 to till date. We retrieved 153 published rickettsial articles on hospital-based studies from India that were purely made on the basis of prevalence and the laboratory parameters viz., Weil-Felix test (WF) and Rapid Immunochromatographic tests (RICT) with reference to the gold standard IFA and ELISA. More epidemiological studies are required for epidemic typhus to know the exact prevalence status of this louse-borne rickettsiosis in India. Currently, there is no confirmed specific inflammatory marker for rickettsial diseases. Moreover, serological cross-reactivity is an important aspect, and it should be investigated in endemic areas, there is also a need to include molecular diagnostic techniques for further confirmation in healthcare settings.

Performance evaluation of a rapid immunochromatographic test kit in detecting bovine mastitis-causing streptococci.

Mastitis causes significant economic losses to the dairy industry due to decreased milk production in infected cows. Identification of mastitis-causing pathogens, such as streptococci, is necessary for selecting an effective antibiotic for treating mastitis. Although bacterial cultivation is widely used for pathogen identification, it requires more than 24 hr to complete. Contrarily, Lateral flow assays are simple, rapid, and inexpensive testing procedures. In this study, the effectiveness of an immunochromatographic test kit for detecting streptococci in milk samples from cows with clinical mastitis was evaluated as an alternative to bacterial cultivation. The performance of the immunochromatographic test kit for detecting mastitis-causing pathogens was compared with that of bacterial cultivation and real-time quantitative polymerase chain reaction (qPCR). The sensitivity and specificity of the immunochromatographic test kit were 0.800 and 0.875, respectively, compared with bacterial cultivation. Additionally, the κ statistic values of the immunochromatographic test kit was 0.667, indicating substantial agreement with the results of bacterial cultivation. Statistically, sensitivity and specificity of the immunochromatographic kit and real-time qPCR did not differ significantly; thus, the immunochromatographic test kit detected mastitis-causing streptococci as effectively as real-time qPCR. Therefore, the immunochromatographic kit is a rapid, inexpensive, and simple method for detecting streptococci and contributes to the timely selection of appropriate antibiotics for treatment and promotes early recovery from mastitis.

Value of the Rapid Immunochromatographic Test for Diagnostic Guidance in the Diagnosis of Pharyngo-Tonsillitis Caused by Group A Hemolytic Beta Streptococcus (Abhs) In Southern Benin

Background: Group A beta-hemolytic streptococcus (ABHS) tonsillitis is an acute disease that is often benign but sometimes serious due to its locoregional and general complications. It is a frequent cause of consultation among general practitioners, ENT and paediatrics. The choice and interest of our work was motivated by the fact that one of the most important acute upper respiratory infections is tonsillitis with the risk of serious complications. This study aims to determine the sensitivity and specificity of RDOT in the diagnosis of ABHA tonsillitis in southern Benin. Methods: This was a prospective analytical and descriptive study carried out at the Vallée de Grâce Medical Clinic (CM VG) and at the Padre Pio Humanitarian Health Center (CSVH), at the Laboratory of Biology and Molecular Typing in Microbiology / UAC, in Benin over 12 months (July 2, 2022 and June 31, 2023). It included patients diagnosed with tonsillitis. All the patients included underwent a RDOT and a cytobacteriological examination of the throat swab (Gold Standard). We studied the following variables: age, sex, months, clinical signs, cytobacteriological examination of the throat swab and TROD results. Results: We recorded 272 tonsillitis cases. 60 cases of group A beta-hemolytic streptococcal (ABHS) tonsillitis are confirmed by bacterial culture. Group A beta-hemolytic streptococcal tonsillitis accounted for 22.06% of tonsillitis. 133 men (49%) and 139 women (51%) were registered. 32 men (53.33%) and 28 women (46.67%) had SBHA tonsillitis. The highest frequency of isolation of group A streptococcus was observed in the age group [0 to 5[years with 27 cases out of 60, i.e. a proportion of 45%. SBHA tonsillitis are more frequent in the months of December and August with the respective proportions of 16.67% and 10%. The SBHA TROD performed showed a Sensitivity of 95%, a Specificity of 94%, a positive predictive value of 92% and a negative predictive value of 98%. Conclusion: Streptococcal etiology represents 22.06% of tonsillitis in Benin. Our results reinforce the interest of RDOT and especially cytobacteriological examination in the etiological diagnosis of tonsillitis.

Comparison of Enzyme Linked Immunosorbent Assay with Immunochromatographic Strip for Screening of Hepatitis B virus Surface Antigen (Hbsag) and Hepatitis C Virus (Anti HCV) Related in a Low Resource Setting

Background: Hepatitis B and C viruses (HBV and HCV) are major causes of liver disease worldwide, with significant prevalence in low-resource settings. Rapid and accurate diagnosis is crucial for effective management and control of these infections. This study compares the efficacy of immunochromatographic tests (ICT) and enzyme-linked immunosorbent assay (ELISA) for the screening of HBV and HCV. Objective: The objective of this study was to evaluate the effectiveness of ICT in comparison with ELISA for the detection of HBsAg and anti-HCV in a semi-urban, low-resource setting. Methods: In this cross-sectional study, 951 blood samples were collected from patients at a tertiary care hospital on the outskirts of Lahore. Samples were tested for HBsAg and anti-HCV using the rapid ICT method (Abbott Bioline HBsAg WB and Abbott Bioline HCV). Positive results were confirmed using ELISA (ANTEC Diagnostic Products, UK). Data analysis was performed using SPSS version 26, focusing on the comparison of sensitivity, specificity, and overall concordance between the two testing methods. Results: Out of 951 individuals tested, the ICT method identified 71 positive cases (7.4%), with 11 (1.2%) for HBsAg and 60 (6.3%) for anti-HCV. ELISA confirmed all 71 positive cases identified by ICT, indicating high concordance between the two methods. The gender distribution was 434 males (45.6%) and 517 females (54.4%), with the majority in the 21-40 year age group (52.2%). Conclusion: ICT is a reliable and cost-effective screening tool for HBsAg and anti-HCV in low-resource settings, with results highly comparable to those obtained from ELISA. Its application could significantly aid in the early detection and management of hepatitis in areas where access to more sophisticated diagnostic facilities is limited.

Evaluation of a rapid immunochromatographic test for the diagnosis of intestinal protozoan infections among patients attending a rural outreach outpatient department in Northern India.

Despite great efforts, intestinal protozoan infections remain a significant healthcare concern worldwide. Although many point-of-care (POC) tests are increasingly being used, microscopic examination of stool specimens remains the mainstay for their diagnosis, especially in resource-limited settings. We assessed the utility of rapid POC tests based on immunochromatography among patients from rural Northern India. A total of 78 patients were enrolled in the study. Out of nine specimens that tested positive for Giardia duodenalis on microscopy, an immunochromatographic test (ICT) could detect only five (55.55%). Entamoeba histolytica/dispar was demonstrated in two specimens on microscopy, both of which were missed by ICT. Its overall sensitivity, specificity, and positive and negative predictive value were 50%, 98.5%, 83.3%, and 93%, respectively. Its performance was considered unsatisfactory. Although ICT-based tests provide a relatively rapid and less labor-intensive alternative, they should be used to supplement and not replace stool microscopy.

Molecular typing of canine parvovirus type 2 by VP2 gene sequencing and restriction fragment length polymorphism in affected dogs from Egypt.

Canine parvovirus-2 (CPV-2) is one of the most common infectious diseases in dogs characterized by severe gastroenteritis, vomiting, and bloody diarrhea. Little information is available about this topic in Egypt, particularly in the Delta region. This study reports the prevalence and molecular analysis of CPV-2 variants collected from El-Gharbia and Kafrelsheikh governorates in the Delta of Egypt. In this study, 320 rectal swabs were collected from infected domestic dogs from two districts in delta Egypt. The samples were investigated by rapid immunochromatographic test and polymerase chain reaction for detection the prevalence of CPV-2 variants. The genetic characterization was performed using restriction fragment length polymorphism (RFLP) analysis and partial VP2 gene sequence. The viral antigen was detected in (264/320, 82.5%) of samples by IC test, while PCR was found more sensitive by detecting (272/320, 85%) positive samples. The RFLP technique using MboII restriction enzyme was successfully used for the differentiation of CPV-2c antigenic variants from CPV-2a/2b strains. Interestingly, the molecular and phylogenetic analysis revealed that both CPV-2a and CPV-2c are circulating in the study area. Deduced amino acid sequence analysis showed changes at residue (N426E) and residue (T440A).: Our results indicated that CPV-2 is prevalent among dogs in Egypt, and therefore further molecular and epidemiological studies of CPV-2 are warranted.

Rapid immunochromatographic test: An evolving tool for diagnosis of scrub typhus

Background: Scrub typhus is prevalent in many districts of South Bengal throughout the year where an average temperature of 20–35°C, which contributes to the spread of Leptotrombidium deliense. However, its diagnosis remains complicated by the lack of readily available and validated assays, the non-specificity of clinical symptoms on admission, and even non-availability of the pathognomonic eschar in most of the cases. Aims and Objectives: This study was carried out to evaluate the rapid immunochromatographic test (RICT) for early detection of scrub typhus for using it as an early diagnostic tool at the field level. Materials and Methods: This cross-sectional study in which 181 serum samples from clinically suspected cases (after excluding dengue, malaria, Japanese encephalitis, and typhoid fever) collected over 13 months were processed for the detection of immunoglobulin M (IgM) antibodies for scrub typhus by enzyme-linked immunosorbent assay (ELISA) and rapid test. Results: Considering IgM ELISA for scrub typhus as the gold standard, the sensitivity, specificity, positive predictive value, and negative predictive value for RICT were found to be 100%, 86.87%, 50%, and 100%, respectively. Conclusion: RICT is a simple, rapid, and reliable assay for diagnosis of scrub typhus, capable of providing accurate results quickly and is highly suitable for field deployment in remote areas with limited medical support.

In house-development of a rapid immunochromatographic test for the detection of Escherichia coli in urine samples

Urinary tract infections (UTIs) are a common health concern. Urine culture is the “gold standard” for UTI diagnosis but takes 48h. Rapid methods like dipstick tests are used as point-of-care tests. However, their sensitivity and specificity are variable. In this work, a rapid immunochromatographic test (IT) for detecting Escherichia coli in urine was developed, and its performance was evaluated in urine samples from patients with suspected UTI. The “universal lateral flow assay kit” was employed using an E. coli capture antibody. One hundred and five (105) urine samples were analyzed using the IT, dipstick test, and urine culture. The sensitivity of the IT was 74.5%, specificity 88.9%, positive predictive value (PPV) 86.3%, and negative predictive value (NPV) 78.7%. The combination of the IT with the dipstick test increases sensitivity to 94.1%, specificity to 66.7%, PPV to 72.7%, and NPV to 92.3%. Using the IT for detecting E. coli in urine could be a valuable technique for UTI screening, showing better specificity and diagnostic precision but lower sensitivity than the dipstick test. Based on these results, we propose that the combined use of both screening techniques would allow a rapid and more precise diagnosis of UTI, rationalizing the indication for empirical antibiotics.

(Un)Reliable detection of menstrual blood in forensic casework - evaluation of the Seratec® PMB test with mock samples.

The identification of the type of body fluid in crime scene evidence may be crucial, so that the efforts are high to reduce the complexity of these analyses and to minimize time and costs. Reliable immunochromatographic rapid tests for specific and sensitive identification of blood, saliva, urine and sperm secretions are already routinely used in forensic genetics. The recently introduced Seratec® PMB test is said to detect not only hemoglobin, but also differentiate menstrual blood from other secretions containing blood (cells) by detecting D-dimers. In our experimental set-up, menstrual blood could be reliably detected in mock forensic samples. Here, the result was independent of sample age and extraction buffer volume. It was also successfully demonstrated that all secretions without blood cells were negative for both, hemoglobin (P) and D-dimer (M). However, several blood cell-containing secretions/tissues comprising blood (injury), nasal blood, postmortem blood and wound crust also demonstrated positive results for D-dimer (M) and were therefore false positives. For blood (injury) and nasal blood, this result was reproduced for different extraction buffer volumes. The results of this study clearly demonstrate that the Seratec® PMB test is neither useful nor suitable for use in forensic genetics because of the great risk of false positive results which can lead to false conclusions, especially in sexual offense or violent acts.