Immunochromatographic Test Kit Research Articles

Epidemiology of an overlapping and parallel infection of Sexually Transmitted Infections among pregnant women in North-east Ethiopia: Its implication for prevention of mother to child transmission.

The burden of parallel and overlapping infections of Sexually Transmitted Infections (STIs), particularly HIV, syphilis, hepatitis B (HBV), and hepatitis C virus (HCV) are disproportionately higher among pregnant women globally, leading to unwanted consequences. These infections pose significant public health challenges as they can be transmitted vertically to the offspring. This study aimed to determine the sero-epidemiological patterns and predictors of STIs (HIV, syphilis, HBV, and HCV) among pregnant women attending antenatal care clinics at ten health facilities in North-eastern Ethiopia. An institution-based multi-center cross-sectional study was conducted from May to November 2022 among 422 pregnant women selected using simple random sampling technique. Semi-structured questionnaire was used to collect socio-demographic characteristics and predictor variables of STIs through face-to-face interviews. Venous blood was collected and it was tested for anti-HIV, HBsAg, anti-HCV, and anti-Treponemal antibodies using immunochromatographic test kits. Multinomial logistic regression analysis was used to identify associated factors of STIs. Variables with an adjusted odds ratio (AOR) and a p-value <0.05 were considered statistically significant. The overall prevalence of STIs was 23.9% (95% CI = 20.08-28.25). The prevalence of parallel infections of HIV, hepatitis B, hepatitis C, and syphilis were 6.4%, 9%, 1.7%, and 6.9%, respectively. The overlapping infections for HIV-HBV was 4% but HIV-HCV overlapping infection wasn't found. Increased age, tattooing, multiple sexual partners, exposure to unsafe sex, and RH status were independent factors of HBV. Likewise, increased age, rural residence, illiteracy, and tattooing were independently associated with HCV. Moreover, rural residence and a history of tattooing were independent predictors for the acquisition of HIV, whereas multiple sexual partners and RH status were found to be significant predictors of syphilis infection among pregnant women. The magnitude of overlapping and parallel STD infections is still continued to be a problem among pregnant women. Moreover, there were overlapping infections of HBV-HIV. Therefore, continuous screening of pregnant women for HIV, syphilis, hepatitis B, and C infections should be performed, and special attention should be given to pregnant women who have co-infections.

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.

Evaluation of a novel triplex immunochromatographic test for rapid simultaneous detection of norovirus, rotavirus, and adenovirus on a single strip test

BackgroundAcute gastroenteritis is one of the major causes of morbidity and mortality in young children worldwide. Among these, rotavirus, norovirus, and adenovirus have been reported as the primary viral pathogens associated with the disease. Rapid diagnosis of viral pathogens is crucial when diarrhea outbreaks occur to ensure the timely administration of appropriate treatment and control measures. MethodsWe evaluated three immunochromatographic test kits designed for the detection of norovirus, rotavirus, and adenovirus in 71 stool specimens collected from children with diarrhea who visited clinics in Japan. The first kit is a triplex immunochromatographic test kit designed for simultaneous detections of norovirus, rotavirus, and adenovirus on a single strip (this kit was referred to as IC-A). The other two immunochromatographic test kits are a dual detection kit for rotavirus and adenovirus, and a single detection kit for norovirus (IC-B). The RT-PCR/PCR was used as the gold standard method. ResultsThe results revealed that both IC-A and IC-B kits exhibited the same level of sensitivity of detection for rotavirus (72.7%) and adenovirus (22.7%), although the detection rate was lower than that of the RT-PCR/PCR method. However, there was a slight difference in the sensitivity of detection for norovirus between IC-A and IC-B, at 86.7% and 93.3%, respectively. The sensitivity of detection for adenovirus of both kits was relatively lower than those of RT-PCR method. This could be due to low viral load of adenovirus in clinical specimens below the detection limit of IC-A and IC-B kits. However, both immunochromatographic test kits (IC-A and IC-B) exhibited 100% specificity for norovirus, rotavirus, and adenovirus. ConclusionsThe triplex immunochromatographic test kit (IC-A) designed for simultaneous detection of norovirus, rotavirus, and adenovirus has been proved to be more practical and convenient than the use of single or dual detection kits with more or less the same sensitivity and specificity of detections.

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.

Etiological Examination of Neonatal Calf Diarrhea Cases Detected in Burdur Region

This work was conducted to determine the prevalence of E. coli, cryptosporidium spp., giardia, rotavirus and coronavirus agents involved in the etiology of diarrhea in cases of neonatal calf diarrhea detected in Burdur Mehmet Akif Ersoy University Animal Hospital Clinics and other farms in the region between 2019-2020 years. The material of the study consisted of 50 neonatal calves with diarrhea. Stool samples were taken from calves with diarrhea and examined with immunochromatographic rapid test kits. One or more enteropathogenic agents were detected in 70% of the calves. In calves with diarrhea, Cryptosporidium spp (42%) and rotavirus (30%) were detected as single and mixed enteropathogen. Mixed enteropathogens (14% Cryptosporidium spp. + Rotavirus, 2% Rotavirus + coronavirus, 2% Cryptosporidium spp. + rotavirus + coronavirus + E. coli K99 + Giardia, 2% Cryptosporidium spp. + coronavirus + E. coli K99 + Giardia) were determined in 20% of calves. As a result, it was concluded that in neonatal calf diarrhea, cryptosporidium and rotavirus factors are common and necessary precautions should be taken in enterprises against these factors.

Evaluation of commercial immunochromatography test kits for diagnosing canine parvovirus

Rapid immunochromatography test (RICT) kits are commonly used for the diagnosis of canine parvovirus (CPV) because of their rapid turnaround time, simplicity, and ease of use. However, the potential for cross-reactivity and low sensitivity can yield false-positive or false-negative results. There are 4 genotypes of CPV. Therefore, evaluating the performance and reliability of RICT kits for CPV detection is essential to ensure accurate diagnosis for appropriate treatment. In this study, we evaluated the performance of commercial RICT kits in the diagnosis of all CPV genotypes. The cross-reactivity of 6 commercial RICT kits was evaluated using 8 dog-related viruses and 4 bacterial strains. The limit of detection (LOD) was measured for the 4 genotypes of CPV and feline panleukopenia virus. The tested kits showed no cross-reactivity with the 8 dog-related viruses or 4 bacteria. Most RICT kits showed strong positive results for CPV-2 variants (CPV-2a, CPV-2b, and CPV-2c). However, the 2 kits produced negative results for CPV-2 or CPV-2b at a titer of 105 FAID50/mL, which may result in inaccurate diagnoses. Therefore, some kits need to improve their LOD by increasing their binding efficiency to detect all CPV genotypes.

Improved diagnostic sensitivity of human strongyloidiasis using point-of-care mixed recombinant antigen-based immunochromatography

Strongyloidiasis is a neglected tropical disease that can cause fatal complications due to hyperinfection and disseminated strongyloidiasis in immunocompromised patients. We used two Strongyloides stercoralis recombinant antigenic proteins, L3NieAg.01 (NIE) and IgG-immunoreactive antigen (SsIR), to develop the recombinant antigen-based immunochromatography test (ICT) kit. We constructed and compared kits using either the NIE (NIE ICT kit) or the SsIR (SsIR ICT kit) antigens and a kit using a mixture of both (NIE-SsIR ICT kit) for detection of anti-Strongyloides IgG antibody in human serum samples. Serum samples from normal healthy individuals (Group I, n = 40), proven strongyloidiasis patients (Group II, n = 100), and those with other parasitic infections (Group III, n = 154) were evaluated. Sensitivity and specificity were 81.0% and 84.0% for the NIE ICT kit, 89.0% and 83.5% for the SsIR ICT kit, and 95.0% and 90.2% for the NIE-SsIR ICT kit, respectively. The NIE-SsIR ICT kit provided the best diagnostic results; it can supplement stool examination for clinical diagnosis and can be used to screen for asymptomatic S. stercoralis infection in people at risk in endemic areas. The NIE-SsIR ICT kit can also be used in large-scale sero-epidemiological investigations in endemic areas without the need for additional facilities or ancillary supplies.

Screening method for tetrodotoxin using immunochromatographic test kit

イムノクロマト法を用いたテトロドトキシン（TTX）検査キットの有効性について検討した。マトリックス非存在下では，0.0125-0.2マウス単位 (MU)/mLの濃度範囲で，テストプレートに形成されるCラインとTラインの強度比（T/C）とTTX濃度の対数の間に強い負の相関が認められ，終濃度約0.06 MU/mLを検出限界として目視によりTTXの有無を判定することができた。一方，マトリックスが存在する場合，その影響を前処理で低減しても偽陽性が出やすかったが，偽陰性は全く認められなかった。

An Innovative Test for the Rapid Detection of Specific IgG Antibodies in Human Whole-Blood for the Diagnosis of Opisthorchis viverrini Infection.

Chronic human liver fluke infections caused by Opisthorchis viverrini and Clonorchis sinensis can last for decades and cause liver and biliary diseases, including life-threatening pathology prior to cholangiocarcinoma (CCA). CCA generally has a poor prognosis. Serological diagnosis can support parasitological examination in diagnosing disease and screening for the risk of CCA. Here, we present an improved and innovative lateral flow immunochromatographic test (ICT) kit that uses whole-blood samples (WBS) rather than serum to diagnose human opisthorchiasis, which also successfully diagnosed human clonorchiasis. This ICT includes a soluble worm extract of O. viverrini adults and colloidal-gold-labeled conjugates of the IgG antibody to evaluate the diagnostic values with simulated WBS (n = 347). Simulated WBS were obtained by the spiking infection sera with red blood cells. The diagnostic sensitivity, specificity, positive and negative predictive values, and accuracy for detecting opisthorchiasis were 95.5%, 87.0%, 80.5%, 97.2%, and 90.1%, respectively. For clonorchiasis, these findings were 85.7%, 87.0%, 53.6%, 97.2%, and 86.8%, respectively. Combined for both diseases, they were 93.2%, 87.0%, 84.0%, 94.6%, and 89.6%, respectively. The ICT kit can possibly replace the ICT platforms for antibody detection in serum samples in field surveys in remote areas where sophisticated equipment is not available.

High Seroprevalence of SARS-CoV-2 in Mwanza, Northwestern Tanzania: A Population-Based Survey.

The transmission of the SARS-CoV-2 virus, which causes COVID-19, has been documented worldwide. However, the evidence of the extent to which transmission has occurred in different countries is still to be established. Understanding the magnitude and distribution of SARS-CoV-2 through seroprevalence studies is important in designing control and preventive strategies in communities. This study investigated the seropositivity of the SARS-CoV-2 virus antibodies in the communities of three different districts in the Mwanza region, Tanzania. A household cross-sectional survey was conducted in September 2021 using the modified African Centre for Disease and Prevention (ACDC) survey protocol. A blood sample was obtained from one member of each of the selected households who consented to take part in the survey. Immunochromatographic rapid test kits were used to detect IgM and IgG SARS-CoV-2 antibodies, followed by descriptive data analysis. Overall, 805 participants were enrolled in the study with a median age of 35 (interquartile range (IQR):27–47) years. The overall SARS-CoV-2 seropositivity was 50.4% (95%CI: 46.9–53.8%). The IgG and IgM seropositivity of the SARS-CoV-2 antibodies was 49.3% and 7.2%, respectively, with 6.1% being both IgG and IgM seropositive. A history of runny nose (aOR: 1.84, 95%CI: 1.03–3.5, p = 0.036), loss of taste (aOR: 1.84, 95%CI: 1.12–4.48, p = 0.023), and living in Ukerewe (aOR: 3.55, 95%CI: 1.68–7.47, p = 0.001) and Magu (aOR: 2.89, 95%CI: 1.34–6.25, p= 0.007) were all independently associated with SARS-CoV-2 IgM seropositivity. Out of the studied factors, living in the Ukerewe district was independently associated with IgG seropositivity (aOR 1.29, CI 1.08–1.54, p = 0.004). Twenty months after the first case of COVID-19 in Tanzania, about half of the studied population in Mwanza was seropositive for SARS-CoV-2.

Hepatitis B virus infection and HBeAg positivity among pregnant women in South West Uganda.

BackgroundHepatitis B virus is a public health burden in Uganda, yet little is known about its epidemiology in pregnancy.ObjectiveThis study aimed at determining the prevalence and associated risk factors of hepatitis B virus infection among pregnant women attending antenatal care at the Kyazanga Health Centre IV in Lwengo District, Uganda.MethodsThis cross-sectional study was conducted from April 2021 to June 2021 and analysed qualitative data that were collected using a structured in-person questionnaire. Aseptically collected blood specimens were screened for hepatitis B virus infection using an immunochromatographic rapid diagnostic test kit. Participants who were positive for the hepatitis B surface antigen (HBsAg) were further screened for hepatitis B envelope antigen (HBeAg) using commercial rapid diagnostic test kits.ResultsOut of 384 pregnant women studied, eight tested positive for HBsAg. This gave a prevalence of 2.1% (95% confidence interval: 1.0% – 4.1%); 5/8 (62.5%) were positive for HBeAg. None of the variables studied were significantly associated with HBsAg positivity among pregnant women.ConclusionHepatitis B viral infection is still a public health challenge in pregnant women with possible risk for vertical transmission to their babies in the study area. We recommend routine screening for hepatitis B virus in pregnancy in addition to strengthening current strategies aimed at controlling and preventing hepatitis B infection spread and transmission.

Detection of canine distemper virus in cerebrospinal fluid, whole blood and mucosal specimens of dogs with distemper using RT-PCR and immunochromatographic assays.

BackgroundAccurate diagnosis of canine distemper (CD), a highly contagious and acute viral disease, cannot be made solely based on clinical signs and haematological findings, but serological and molecular methods compatible with clinical signs are also required. The type of sample and method of tissue sampling are also very important. Sometimes in chronic cases, the canine distemper virus (CDV) may not be detected in blood and conjunctival specimens but can be detected in cerebrospinal fluid (CSF).ObjectivesThe aim of this study was to evaluate and compare the suitability of CSF samples with whole blood and conjunctival samples in the detection of CDV.MethodsThe CDV was detected in CSF, whole blood and mucosal specimens in 20 dogs with obvious neurological with or without systemic signs congruous with CD by RT‐PCR and rapid immunochromatographic (IC) antigen test kit assays.ResultsRapid kit results were positive for mucosal swabs in 10 cases (50%) and for CSF in 17 cases (85%); RT‐PCR results from whole blood were positive in 11 cases (55%) and from CSF in 16 cases (80%).ConclusionsOur results revealed that dogs with neurological signs showing simultaneous or recent systemic symptoms, whole blood, CSF and mucosal swabs are suitable for the diagnosis of CDV by RT‐PCR and rapid IC antigen test kits, but dogs with neurological symptoms that are systematically asymptomatic or have had systemic signs for a long time, whole blood and mucosal swabs are not good samples while CSF is a good one.

Detection of Epsilon Toxin in Goats Affected with Acute Enterotoxaemia and the Haemato-biochemical Alterations

Background: The principal cause of caprine enterotoxaemia is Clostridium perfringens type D which is normally resident in digestive tract and soil in fairly low numbers. Overeating of carbohydrate rich diet or sudden change in diet leads to proliferation of the bacteria and release of lethal toxins in intestine of the host. It causes sudden death in affected ruminants of different ages, though lambs of 3 to 10 weeks of age are highly susceptible. It causes sever enterocolitis with or without haemorrhagic diarrhoea and encephalitis resulting in acute nervous signs with a fatal outcome. Methods: Thirty goats of different breeds under 1-4 years age and both sex were presented to Veterinary Clinical Complex, Veterinary College and Research Institute, Tirunelveli with the neurological signs and diarrhoea were the subject of study. Faecal samples were collected from all these cases and tested on spot by immunochromatographic test kit to identify the Clostridium perfringens Epsilon toxin (ETX). Blood and serum samples were collected and subjected haemato-biochemical analysis. Peripheral blood smears were also collected to rule out haemoprotozoan infections in all the cases. Result: Epsilon toxin was detected from faecal samples of 36.6 per cent of the goats. Diarrhoea was the most consistent finding which is due to enterocolitis in acute cases. A significant increase (P less than 0.001) in the neutrophils and leukocytes count and a significant increase (P less than 0.05) in the levels of BUN, creatinine, glucose, ALP and chloride levels were observed. All the cases were treated with sulphadimidine at recommended dose with supportive therapy but no response was reported due to the fatal encephalitis.

Serum Calprotectin Levels in Dogs with Diarrhea

Background: Diarrhea induced by infectious factors may lead to significant health problems in dogs. Canine parvovirus (CPV), canine coronavirus (CCV), canine distemper virus (CDV), Giardia spp., Escherichia coli (E. coli), and Salmonella spp. are the important infectious agents that may induce diarrhea in dogs. The present study aimed to investigate the effect of CPV, CCV, CDV, Giardia spp., E. coli, and Salmonella spp. infections on the change in serum calprotectin (Calp) concentration.Materials, Methods &amp; Results: A total of 30 dogs were enrolled in the study. The study dogs were divided into 3 groups. Healthy animals as confirmed by clinical examination and animals negative for the specified pathogens were placed in Group 1. Animals infected by one or more agents, including CPV, CCV, CDV, and Giardia spp., but negative for E. coli or Salmonella spp. were placed in Group 2. Finally, animals positive for E. coli or Salmonella spp. and infected or not infected by one or more agents, including CPV, CCV, CDV, and Giardia spp., were placed in Group 3. Stool samples and rectal and conjunctival swab samples were collected to investigate the etiologic agents that induced diarrhea. Blood samples were collected through vena cephalica antebrachii for hematological and biochemical examinations. The samples were obtained via routine clinical examinations at the Prof. Dr. Servet SEKİN outpatient clinic at Dicle University Veterinary Faculty. CPV, CCV, CDV, and Giardia spp. diagnoses were made based on immunochromatographic test kits. The bacteriological analysis of stool samples were used to diagnose E. coli and Salmonella spp. infection. Serum Calp concentrations were measured by Enzyme-Linked Immunosorbent Assay (ELISA). The analysis of swab and stool samples by immunochromatographic rapid diagnosis kits and microbiological methods showed that 5 animals were infected with CPV, 10 with CCV, 6 with CDV, 3 with Giardia spp., 12 with E. coli, and 2 with none of the specified agents. Total leukocyte count (WBC), lymphocyte (Lym) (%), and granulocyte (Gra) (%) values were higher in the diarrheal dogs when compared with the control group. In the biochemical examination of serum samples, TBIL and P levels were higher and Na levels were lower in Group 3 when compared to the control group (P = 0.025, 0.024, and 0.018, respectively). TP and Alb values were lower in Group 2 compared to Groups 1 and 3 (P = 0.001 and 0.019 for TP, P = 0.000 and 0.001 for Alb, respectively). There was a statistically significant difference in CK levels between Group 1 and Group 2 (P = 0.013). Serum Calp level was higher in the E. coli infected group (Group 3) compared to the other groups, no significant differences were noted between the groups (P &gt; 0.05).Discussion: In conclusion, to the best of authors knowledge, this study is the first to evaluate serum Calp levels in dogs with diarrhea induced by viral, bacterial, and protozoan infections. The Calp level was higher in the sick dogs that were infected by at least one agent, including CPV, CCV, CDV, and Giardia spp., and were at the same time E. coli positive when compared with the control group and the group without E. coli infections. It was concluded that new studies could be useful to reveal the diagnostic importance of serum Calp concentration in dogs with diarrhea and that these results may contribute to future studies in this area.Keywords: calprotectin, diarrhea, dog, serum.

GroEL Chaperonin-Based Assay for Early Diagnosis of Scrub Typhus.

A point-of-care diagnostic for early and rapid diagnosis of scrub typhus caused by Orientia tsutsugamushi is required for prompt and proper treatment of patients presenting with undifferentiated febrile illnesses. In this study, an immunochromatographic antigen detection test kit (ICT AgTK) that targets the highly conserved O. tsutsugamushi 60 kDa GroEL chaperonin (heat shock protein 60) was developed. E. coli-derived recombinant GroEL expressed from DNA coding for the consensus sequence of 32 GroEL gene sequences extracted from the GenBank database was used to immunize rabbits and mice. Rabbit polyclonal antibodies (pAb) were used for preparing a gold-pAb conjugate, and the rGroEL-specific mouse monoclonal antibody was used as the antigen detection reagent at the ICT test line. In-house validation revealed that the ICT AgTK gave 85, 100 and 95% diagnostic sensitivity, specificity and accuracy, respectively, compared to the combined clinical features and standard IFA when tested on 40 frozen serum samples. The test kits correctly identified 10 scrub typhus samples out of 15 fresh plasma/buffy coat samples of patients with febrile illnesses. For independent laboratory validation, the ICT AgTK was sent to one provincial hospital. The ICT AgTK utilized by the hospital medical technologist correctly identified six scrub typhus samples out of 20 serum samples of patients with fever, as confirmed by specific IgM/IgG detection by IFA. The ICT AgTK is easy to perform with rapid turn-around time. It has the potential to be used as an important tool for on-site and early scrub typhus diagnosis by allowing testing of freshly collected samples (serum, plasma or buffy coat), especially in resource-limited healthcare settings.

Occurrence of Dirofilaria immitis in cats from the Aegean region in Turkey

Dirofilaria immitis (D. immitis) is a vector-mediated zoonotic parasite that can cause significant cardiopulmonary problems in humans and animals. Although dogs are the main hosts of the parasite, in recent years, its importance has been increasing in cats, especially in endemic areas. This study aims to determine the occurrence of D. immitis infection in cats in the Aegean region, Turkey. The animal material of the study consisted of 200 cats of different breeds and ages (at least 1 year old) and both sexes. The cats were tested for D. immitis and Feline Immunodeficiency Virus (FIV) antibodies and Feline Leukaemia Virus (FeLV) antigen with commercial immuno-chromatographic test kits. In addition, blood samples were examined for microfilariae using a direct blood smear technique and the modified Knott’s test. Out of 200 cats, one (0.5%) cat was seropositive for D. immitis, 15 (7.5%) cats were seropositive for FIV, and one (0.5%) cat was seropositive for FeLV. One cat seropositive for D. immitis was showing signs of active heartworm infection. All samples were found negative for microfilariae by direct blood examination and the modified Knott’s method. This study is the first report of the occurrence of D. immitis in the Aegean Region, with a seroprevalence of 0.5%. Also, it provides evidence that cats in the Aegean region are at risk of becoming infected with D. immitis.

Rapid assessment of Opisthorchis viverrini IgG antibody in serum: A potential diagnostic biomarker to predict risk of cholangiocarcinoma in regions endemic for opisthorchiasis

BackgroundOpisthorchiasis is caused by an infection with fish-borne liver flukes of the genus Opisthorchis. Opisthorchiasis frequently leads to chronic inflammation in the biliary tract and is classified as a group 1 biological carcinogen by the International Agency for Research on Cancer: a definitive risk for cholangiocarcinoma (CCA). MethodsWe used the rapid immunochromatographic test (ICT) to detect anti-Opisthorchis viverrini IgG and IgG4 subclass antibodies in sera of patients with CCA. The ICT kits were developed based on soluble antigens excreted and secreted by O. viverrini adult worms. ResultsICT indicated sera was positive for IgG and IgG4 antibodies, respectively, in 22 (61.1%) and 15 (41.6%) participants of the 36 study participants diagnosed with CCA (P > 0.05). Our study also included groups with other cancers and with liver cirrhosis, where the IgG ICT and IgG4 ICT kits were 27.7% (13/47) and 25.5% (12/47) positive, respectively (P > 0.05). Neither total the IgG ICT nor the IgG4 ICT yielded positive results in a control group of 20 healthy participants. Moreover, the percentage positivity rate using the ICT for total IgG between the CCA group and the other cancers and liver cirrhosis group was significantly different (P < 0.05). By contrast, no significant difference between these groups was apparent in the ICT for IgG4 antibody. The CCA group was 6.53 times more likely to have positive anti–O. viverrini IgG antibody (odds ratio 6.53, P < 0.001) and 3.27 times more likely to have positive anti–O. viverrini IgG4 antibody (odds ratio 3.27, P = 0.010) than the non-CCA group. ConclusionThis information is of potential value for the development of a diagnostic biomarker to predict risk for O. viverrini infection-associated CCA.

Prevalence of Clinical Malaria in District Khairpur Mir’s Sindh, Pakistan

Aim: To evaluate the frequency of malarial parasites in different seasons at district Khairpur Mir’s, Sindh, Pakistan. Study Design: A prospective study. Place and study duration: This study was conducted at Post Graduate Diagnostic and Research Laboratory (PGRDL), Institute of Microbiology Shah Abdul Latif University (SALU), Khairpur and Bismillah Medical Center (BMC), Khairpur, from 01-Jan-2021 to 30-June-2021. Methodology: Febrile patient of all ages and both sexes were included, pre-diagnosis of malarial parasites using the Immunochromatographic test (ICT) kits, the blood samples were taken by venepuncture procedure after collecting samples in Ethylene diamine tetra acetic acid (EDTA) tubes for ICT and blood slides for microscopy. All results were entered and analyzed in SPSS 24 version. Continuous variables were presented as mean and SD and qualitative variables were presented as frequency/percentages. Probability value ≤0.05 was considered as significant. Results: In this study overall 7332 febrile patients with means age 37.8±22.54, from which male 3476 (47%) and majority of 3856 (53%) were female patients. Whereas 1908 (26%) patients were suffered from the active state of malaria and majority of 5424 (74%) population suffered from non-malarial diseases having vague symptoms including the chills, fever, body aches, abdominal cramps. The positive cases of Plasmodium vivax 1534 (20.92%) Plasmodium falciparum 316 (4%) whereas mixed cases are 58 (1%). Conclusion: The malarial parasites were observed predominantly with type of P. Vivax followed by P. falciparum and mix cases. Therefore dire need to take preventive measures by controlling the spread of plasmodium species in month of March onward till June. Keywords: Malarial Parasites (MP), Febrile Patients, Immunochromatographic Test (ICT).

A novel gold nanosensor and ‘Blockade-of-Binding’ based immunochromatographic rapid antigen test kit for Zika virus

The present study relates to the development of an immunochromatographic, antigen test kit for rapid detection of Zika virus using stable gold nanoparticles (AuNPs). Herein, we have developed an alternate viral NS1 antigen (Ag) based rapid test that uses monoclonal antibody (Mab) pairs to detect ZIKV without cross-reactivity in 5 min. To mask the non-specific results, the “Blockade of Binding' assay was employed in lateral flow kits. Also, for better performance of kits, a sample pad having an absorption capacity of 20 µl/cm2 was utilized. Herein, AuNPs-ZIKA NS1 Mab is capable of binding to specific sites of the Zika antigen. Anti-DENV-MAb is capable of binding to a specific site for Dengue, anti-CHKV-Mab is capable of binding to a specific site for Chikungunya, ZIKA NS1 Mab is capable of binding to anti-ZIKA NS1 Mab and GAM Mab is capable of binding to leftover anti-ZIKA NS1 MAb. The kit was found to be stable for 12 months while its specificity and sensitivity were found to be 95.83% and 100%, respectively.

THE PREVALENCE OF CHRONIC HEPATITIS C IN PATIENTS PRESENTING WITH VAGUE RHEUMATIC SYMPTOMS IN A LOCAL COMMUNITY IN DISTRICT PUNJAB, PAKISTAN

Objective:&#x0D; This study has been conducted to highlight the prevalence of chronic hepatitis in patients presenting with vague symptoms such as generalized body aches and weakness.&#x0D; Methods:&#x0D; A cross-sectional study was conducted in Tehsil Headquarter Hospital Sharaqpur, Sheikhupura, Pakistan. During a period of two and a half months, 751 patients presenting to general outdoor department with vague symptoms were selected through random sampling. They were tested for Anti Hepatitis C Virus Antibodies, Hepatitis B Surface Antigen, and Anti Human Immunodeficiency Virus Antibodies with Rapid Immunochromatographic Test Kits. Statistical analysis was performed using IBM SPSS Statistics version 23.&#x0D; Results:&#x0D; Of the 751 randomly selected patients, 28 were eliminated due to missing data. Out of 723 included participants, 36% were male and 64% were female with a mean age of 53.11±11.2 years. 26.42% people were screened positive for Hepatitis C (N=191). 1.7% people were screened positive for Hepatitis B (N=13). Whereas only 0.12% (N=1) were HIV positive.&#x0D; Conclusion:&#x0D; This study established that Chronic Hepatitis often presents with mild and vague symptoms. Also, the prevalence of HCV is significantly higher in our population in spite of the ongoing hepatitis control programs and is rising in contrast to the global decline.