Enzyme-linked Immunosorbent Assay Positive Samples Research Articles

Evaluation of Aspergillus Galactomannan Lateral Flow Assay on Serum and Bronchoalveolar Lavage Specimens

Background: Detection of biomarkers, such as galactomannan (GM), has proven to be of great significance in early recognition of invasive aspergillosis (IA). The aim of our study was to evaluate the lateral flow assay (LFA) for the detection of GM on serum and bronchoalveolar lavage (BAL) samples previously proven positive by enzyme-linked immunosorbent assay (ELISA). Methods: The study was performed on serum and BAL samples obtained from patients with suspected IA in the period from February 2019 to January 2020, which were previously GM positive by ELISA (Platelia Aspergillus Ag, Biorad, Hercules, USA). Samples were then tested by LFA (Aspergillus Galactomannan LFA, IMMY, Oklahoma, USA) with test line intensity visually read as 1+, 2+, 3+, or 4+. Results: A total of 45 GM ELISA positive serum and/or BAL samples were obtained from 41 patients; 25 (55,6 %) were BAL and 20 (44,4 %) serum samples. LFA showed a positive result in 39 out of 45 (86,7%) GM ELISA positive samples; 22/25 (88.0 %) BAL samples and 17/20 (85.0 %) serum samples tested positive. In BAL samples, low intensity test line of 1+ was significantly more frequent in GM ELISA positive samples with optical density index (ODI) < 1.0 (p=0.0002). Three serum samples with high GM ELISA ODI (>4.0) had low intensity line of 1+ when tested with LFA. Conclusions: Results obtained by LFA are comparable to GM ELISA. Since low intensity lines were found in serum samples with high ODI, this potentially makes BAL a superior sample for LFA, at least when visual and not automated reading is done.

Common hepatitis B virus genotypes among blood donors in Lagos, Nigeria.

Hepatitis B virus (HBV) infection is one of the public health diseases causing global health problems. It is a leading cause of cirrhosis and hepatocellular carcinoma. Blood transfusion is a major route of its transmission and screening of blood is suboptimal in our environment. Occult HBV infection describes the presence of HBV DNA in blood or liver tissue in patients who are hepatitis B surface antigen (HBsAg) seronegative. This study documented the common genotypes of HBV a blood-borne infection in the population of blood donors in Lagos. This was a cross-sectional study carried out at the blood donor clinics of ten Lagos State Government-owned hospitals in Lagos State. A total of 1400 participants were recruited consecutively from November 2020 to June 2021. All participants' samples were screened using Diaspot Rapid Test Kit (RTK) and Dialabenzyme enzyme-linked immunosorbent assay (ELISA) kit. Furthermore, some of the plasma samples were used for HBV DNA extraction and genotyping using the real time-polymerase chain reaction. Statistical analysis was carried out using the Statistical Package for the Social Sciences (SPSS) software version 26 and P value was considered significant at ≤0.05. The sero-prevalence of HBsAg using RTK and ELISA was 19.9% and 22.4%, respectively. The prevalence of occult HBV infection was 5.2%. A total of 278 and 313 HBsAg RTK and ELISA positive samples were obtained, respectively. HBV genotype result had A (46.6%) as the most prevalent followed closely by B (44.7%), E (23.8%), D (20.9%) and C (11.2%). HBV infection has a high prevalence among blood donors. ELISA is a more sensitive screening tool and its use should be advocated nationally. HBV genotype A is the most prevalent genotype from our study.

Seroprevalence and characterisation of herpes simplex virus from human immunodeficiency virus in samples collected from two provinces in South Africa: a retrospective study.

Background:Herpes simplex virus (HSV) is a widely distributed human pathogen that is known for its ulcerative lesions at the infection site. HSV can cause persistent infection in the host that is often followed by a period of latency within the neurons. Considering the high rate of HIV infection in South Africa, it is important to assess the seroprevalence of HSV with a focus to determine the epidemiological association between HSV-DNA and HIV-1 in the population. Methods:A total of 44 sera samples were screened for HSV and HIV-1 using the highly sensitive enzyme-linked immunosorbent assay (ELISA). The ELISA positive samples were characterized using polymerase chain reaction (PCR) to confirm the positivity of both viruses and to further differentiate HSV into HSV-1 and -2. Thereafter, the samples were analysed for relatedness using phylogenetic analysis. Results:Of the 44 samples, 36 (81.8%) were positive for HIV-1, while 35 (79.5%) were positive for HSV when screened with ELISA kits. The PCR results, with the use of type specific primers, showed that 4/35 (11.4%) samples were specific for HSV-1 while 30/35 (85.7%) were specific for HSV-2. Statistical analysis performed using the chi-squared goodness-of-fit test showed that there is a significant relationship between HSV-2 and HIV-1 transmission. Conclusions:There is a significant positive association between HSV-2 and HIV-1 in the study population. Our study shows that some of the HSV-2 isolates are not related to the clinical isolate SD90e from South Africa, suggesting diversity in HSV-2 viral transmission.

Investigation of Group A Rotavirus G10, G12 Genotypes Emerging in Patients with Acute Gastroenteritis in a Tertiary Care Hospital

Rotaviruses are the most common cause of viral gastroenteritis with the highest mortality and morbidity rates in children aged 0-5 years. The aim of this study was to determine the frequency of rotavirus infection in patients whose stool samples were sent to microbiology laboratory to investigate the etiology of diarrhea, to investigate the rotavirus genotypes that are common in our region and G10, G12 genotypes that have recently become common in the world. Fecal samples of 476 patients aged between 0-92 years who applied between November 2016 and February 2018 were studied via immunochromatographic rapid test and enzyme-linked immunosorbent assay (ELISA) methods. ELISA positive samples were studied by nested reverse transcriptase chain reaction (RT-PCR) and genotyped by agarose gel electrophoresis. Rotavirus was found positive in 18.3% and 17% of stool samples by immunochromatographic test and ELISA, respectively. All ELISA positive samples were also detected as positive by RT-PCR. 18.5% of female patients and 15.7% of male patients were found to be positive and rotavirus positivity was not statistically significant between genders. The frequency of rotavirus in different age groups was 23.5% (6-12 years), 17.3% (13-24 months) and 16% (25-36 months). It was determined that rotavirus cases were most common in the spring. G1, G2, G3, G4, G9, G10, and G12 were detected in 37%, 7.4%, 16.1%, 6.2%, 9.9%, 2.5%, 26% of the samples, respectively. G12 was the most common genotype after G1. The most common G and P genotype combination was G1P[8] (17.2%). This was followed by G12P[8] (11.11%) and G3P[8] (11.11%). P[8] (53%) was found to be the dominant P genotype. In this study, it was observed that rotavirus, which is the cause of childhood diarrhea, can also be encountered in advanced ages and even new genotypes that infect humans worldwide may also be the causative agents. Therefore, we concluded that it is important to investigate new genotypes such as G10 and G12 in molecular epidemiological studies.

Seroprevalence and characterisation of herpes simplex virus from human immunodeficiency virus in samples collected from two provinces in South Africa: a retrospective study

Background: Herpes simplex virus (HSV) is a widely distributed human pathogen that is known for its ulcerative lesions at the infection site. HSV can cause persistent infection in the host that is often followed by a period of latency within the neurons. Considering the high rate of HIV infection in South Africa, it is important to assess the seroprevalence of HSV with a focus to determine the epidemiological association between HSV-DNA and HIV-1 in the population. Methods: A total of 44 sera samples were screened for HSV and HIV-1 using the highly sensitive enzyme-linked immunosorbent assay (ELISA). The ELISA positive samples were characterized using polymerase chain reaction (PCR) to confirm the positivity of both viruses and to further differentiate HSV into HSV-1 and -2. Thereafter, the samples were analysed for relatedness using phylogenetic analysis. Results: Of the 44 samples, 36 (81.8%) were positive for HIV-1, while 35 (79.5%) were positive for HSV when screened with ELISA kits. The PCR results, with the use of type specific primers, showed that 4/35 (11.4%) samples were specific for HSV-1 while 30/35 (85.7%) were specific for HSV-2. Statistical analysis performed using the chi-squared goodness-of-fit test showed that there is a significant relationship between HSV-2 and HIV-1 transmission. Conclusions:There is a significant relationship between HSV-2 and HIV-1 in the study population. Our study shows that some of the HSV-2 isolates are not related to the clinical isolate SD90e from South Africa, suggesting diversity in HSV-2 viral transmission.

Seroprevalence and risk factors of Toxoplasma gondii infection among veterinary personnel and abattoir workers in Central India.

Toxoplasmosis, caused by the protozoan parasite Toxoplasma gondii, is an important zoonotic infection. Veterinary personnel and abattoir workers are considered to be at a high risk of T. gondii infection owing to their occupational exposure. However, the association of T. gondii infection with occupational exposure to animals has not been determined in India. Hence, we analysed 139 and 126 blood samples of veterinary personnel and abattoir workers, respectively, for anti-T. gondii antibodies using enzyme-linked immunosorbent assay (ELISA), modified agglutination test (MAT) and indirect fluorescent antibody test (IFAT). The association of seroprevalence with sociodemographic profiles, work activities and dietary habits was determined in the study population. MAT, ELISA and IFAT results demonstrated nearly 46%, 48% and 47% seropositivity, respectively. MAT (kappa=0.924) and IFAT (kappa=0.962) results showed good agreement with ELISA results. Of the ELISA positive samples, 46% was copositive for IgG antibody, 1.5% for IgM antibody and 1.5% for both IgG and IgM antibodies. High IgG avidity was observed only in IgG+ IgM- and IgG+ IgM+ samples and not in IgM+ IgG- samples, indicating chronic T. gondii infection in most of the cases. Furthermore, multivariate analysis revealed that T. gondii seropositivity was associated with age>30years (odds ration [OR]=1.992), cat at home (OR=1.991), not wearing gloves (OR=1.886), not wearing safety glasses (OR=1.985) and contact with soil (OR=1.695). These findings support the presence of a potentially significant association between T. gondii seropositivity and occupational exposure to animals.

Seroprevalence and characterisation of herpes simplex virus from human immunodeficiency virus in samples collected from two provinces in South Africa: a retrospective study

Background: Herpes simplex virus (HSV) is a widely distributed human pathogen that is known for its ulcerative lesions at the infection site. HSV can cause persistent infection in the host that is often followed by a period of latency within the neurons. Considering the high rate of HIV infection in South Africa, it is important to assess the seroprevalence of HSV with a focus to determine the epidemiological association between HSV-DNA and HIV-1 in the population. Methods: A total of 44 sera samples were screened for HSV and HIV-1 using the highly sensitive enzyme-linked immunosorbent assay (ELISA). The ELISA positive samples were characterized using polymerase chain reaction (PCR) to confirm the positivity of both viruses and to further differentiate HSV into HSV-1 and -2. Thereafter, the samples were analysed for relatedness using phylogenetic analysis. Results: Of the 44 samples, 36 (81.8%) were positive for HIV-1, while 35 (79.5%) were positive for HSV when screened with ELISA kits. The PCR results, with the use of type specific primers, showed that 4/35 (11.4%) samples were specific for HSV-1 while 30/35 (85.7%) were specific for HSV-2. Statistical analysis performed using the chi-squared goodness-of-fit test showed that there is a significant relationship between HSV-2 and HIV-1 transmission. Conclusions: The prevalence of HSV in the population is high with an increased HSV-2 infection in women. Our study shows that some of the HSV-2 isolates are not related to the clinical isolate SD90e from South Africa, suggesting diversity in HSV-2 viral transmission.

Seroprevalence and characterisation of herpes simplex virus from human immunodeficiency virus in samples collected from two provinces in South Africa: a retrospective study

Background: Herpes simplex virus (HSV) is a widely distributed human pathogen that is known for its ulcerative lesions at the infection site. HSV can cause persistent infection in the host that is often followed by a period of latency within the neurons. Considering the high rate of HIV infection in South Africa, it is important to assess the seroprevalence of HSV with a focus to determine the epidemiological association between HSV-DNA and HIV-1 in the population. Methods: A total of 44 sera samples were screened for HSV and HIV-1 using the highly sensitive enzyme-linked immunosorbent assay (ELISA). The ELISA positive samples were characterized using polymerase chain reaction (PCR) to confirm the positivity of both viruses and to further differentiate HSV into HSV-1 and -2. Thereafter, the samples were analysed for relatedness using phylogenetic analysis. Results: Of the 44 samples, 36 (81.8%) were positive for HIV-1, while 35 (79.5%) were positive for HSV when screened with ELISA kits. The PCR results, with the use of type specific primers, showed that 4/35 (11.4%) samples were specific for HSV-1 while 30/35 (85.7%) were specific for HSV-2. Statistical analysis performed using the chi-squared goodness-of-fit test showed that there is a significant relationship between HSV-2 and HIV-1 transmission. Conclusions: There is a significant positive association between HSV-2 and HIV-1 in the study population. Our study shows that some of the HSV-2 isolates are not related to the clinical isolate SD90e from South Africa, suggesting diversity in HSV-2 viral transmission.

Seroprevalence and characterisation of herpes simplex virus from human immunodeficiency virus in samples collected from the North-West and KwaZulu-Natal Provinces: a retrospective study.

Background: Herpes simplex viruses (HSVs) are highly pervasive and show a strongsynergistic interaction with human immunodeficiency virus (HIV). High prevalence of HSV type 1 (HSV-1) has been reported in Africa with a prevalence rate of 20-80% in women and 10-50% in men. Studies on the prevalence of HSV in South Africa are few considering the rate of HIV infection in the country. Our focus was to determine the molecular prevalence of HSV-DNA in HIV-1 sera. Methods: In total, 44 convenience samples were screened for HSV and HIV-1 using the highly sensitive enzyme-linked immunosorbent assay (ELISA). The ELISA positive samples were characterized using polymerase chain reaction (PCR) to confirm the positivity of both viruses and to further differentiate HSV into HSV-1 and -2. Thereafter, the samples were analysed for relatedness using phylogenetic analysis. Results: Of 44 samples, 36 (81.8%) were positive for HIV-1, while 35 (79.5%) were positive for HSV when screened with ELISA kits. The results of PCR with type specific primers showed that 4/35 (11.4%) samples were specific for HSV-1 while 30/35 (85.7%) were specific for HSV-2. Statistical analysis performed using chi-squared goodness-of-fit test showed that there is a significant relationship between HSV-2 and HIV-1 transmission. Conclusions: High prevalence of HSV-2 recorded in HIV-1 sera corroborate with similar studies conducted within different cohorts in the continent. SPSS Pearson's chi-squared test established that there is a significant relationship between HSV-2 and HIV-1 transmission.

Efficiency of molecular detection of Toxoplasma gondii contrasting to serological method.

Toxoplasmosis infection of the mother during pregnancy is linked to transplacental transmission to the fetus. Toxoplasma gondii infection is difficult to diagnose and necessitates a comprehensive examination that includes laboratory and clinical testing. This study used RT-PCR using B1 gene based on paired blood samples and clinical data, for the detection of Toxoplasma infections in miscarriage women, this technique was compared to enzyme-linked immunosorbent assay (ELISA). There are a total of 125 miscarriage women at Al-Muthanna Hospital. Anti-Toxoplasma antibodies, IgG (latent infection) and IgM (recently acquired infection), were positive in 50/125 cases, with 25.6% IgG, 8.8% IgM, and 5.6% for IgM and IgG. In addition, women who have had two miscarriages have a high seropositivity rate. Furthermore, the development of molecular technologies to amplify parasite nucleic acids has enhanced toxoplasmosis diagnosis. The cycle threshold values Ct, which indicate the target gene's quantity, were calculated. With DNA from 500 T. gondii tachyzoites, a Ct of 25-28 was usually obtained. Present results show that the assay's reproducibility was found in 50 which considered (40%) from total miscarriage women of ELISA positive samples while 28/125 samples (22.4%) will positively in molecular detection and 56% from serological test was positive in molecular test. A move forward was the invention of diagnostic methods that used ELISA IgG and IgM followed by an RT-PCR methods in women were the most effective in detecting recent and reactive Toxoplasma gondii infection.

Detection of Hepatitis C Virus Antibodies among Healthy Blood Donors at a Tertiary Hospital in Nigeria

Background: Most blood transfusion centres in Nigeria including the University of Abuja Teaching Hospital (UATH), Gwagwalada, Abuja use rapid test kit for screening blood donors. Though it is simple, robust with speedy results, it has some limits. This cross-sectional study determined the prevalence of HCV antibodies by the currently used rapid detection assay among blood donors tested positive by ELISA, describe the HCV risk factors and socio-demographic characters among the blood donor participants. Methods: Blood samples collected from 365 blood donors attending UATH blood bank were screened for HCV antibodies using Enzyme-linked Immunosorbent Assay (ELISA) kit (CTK Biotech, USA). Positive samples were retested using Aria rapid test kit. Structured questionnaires were used to collate subjects’ socio-demographic data and risk factors of infection. Results: Out of 365 blood donors tested, 55 (15.1%) were positive for HCV antibodies. Of the 55 ELISA positive samples, 9 (16.4%) samples were negative by Aria rapid test kit. Sociodemographic data showed that female had a higher prevalence (16.3%) than the male (14.9%) p= 0.81. The age group with the highest rate of infection was 31-45 years p. Conclusion: The study revealed the superiority of ELISA over Aria rapid test kit in the detection of HCV antibodies. The missed positive cases by the currently used rapid test could pose a risk to blood transfusion safety and necessitates for preventive measures to be intensified, as blood donors with high risk factors be deferred from blood donation to reduce HCV transfusion risk in Nigeria.

First Full Length Genome Sequence of Bean Common Mosaic Necrosis Virus (BCMNV) Isolated from Common Bean in Western Kenya

Bean common mosaic necrosis virus (BCMNV) is one of the most common and most destructive viruses of common bean and can cause a yield loss as high as 100%. The common bean (Phaseolus vulgaris L) is an important legume crop for food and cash in Kenya. In Kenya, there is inadequate documentation on the strains of the virus infecting common bean. This information is crucial in devising control measures. This study therefore, sought to characterize BCMNV isolates from western Kenya. Leafy samples showing virus-like symptoms were collected and analysed by Enzyme linked immunosorbent Assay (ELISA) and or next generation sequencing (NGS). Extraction of total RNA from ELISA positive samples was done using RNeasy Plant Mini Kit and NGS carried out following Illumina protocol to determine diversity of the virus. NGS data was trimmed and the sequence reads assembled into contigs, which were analyzed against virus sequence database. Phylogenetic analyses and comparisons were performed using MEGA7 program. The first complete genome sequence of Bean common mosaic virus (BCMNV) is reported from a Kenyan isolate. NGS technology revealed full-length sequence of BCMNV from an isolate BG 12 from Bungoma County with a genome of 9584 nt in length. Phylogenetic analysis of full-length sequences available through the Genbank clustered the isolate with the Tanzanian isolate strain TN-1 and two USA isolates, TN1a and NL-3K.

West Nile virus - a new infection in the Slovak Republic?

The aim of the study was to evaluate the seroprevalence of West Nile virus (WNV) among the variable population of Eastern Slovakia. A serologic survey was conducted using 464 serum samples. The basic demographic, epidemiologic and clinical information was obtained for each serum sample at the time of specimen collection. The presence of antibodies against WNV was investigated using a commercial enzyme-linked immunosorbent assay (ELISA). All the ELISA positive samples were further analysed by a neutralization test with WNV and Usutu virus. Three serum samples (0.65%) from the participants (N = 464) were considered positive for antibodies to WNV. A 29-year-old female was repeatedly exposed to mosquito bites working as a shepherdess and participating in many outdoor activities. Two other females (61 and 76 years old) were treated at the Department of Neurology due to monoparesis of the upper extremity, vertigo; both had a significant epidemiological history with frequent tick and mosquito bites and stay in an endemic region. Although there was no evidence of WNV infection in the Slovak Republic, the epidemiological situation in the neighbouring countries warrants vigilance and appropriate measures, including the introduction of specific diagnostic tools into clinical practice. The constant monitoring of birds and mosquitoes also seems necessary.

Serological evidence of Brucella infections in dairy cattle in Haryana, India

ABSTRACTBackground: In India, milk production is important for both the economy and the provision of nutritious food. However, the productivity of the livestock is affected by circulating infectious diseases, and some zoonotic diseases, such as brucellosis, may cause a heavy impact on the farm as the disease cause abortions and reproductive failures in bovines, with chronic febrile illness in humans.Methods: 249 dairy farms in the state of Haryana, India, were interviewed, and collected raw milk from 81 were analyzed for antibodies towards Brucella abortus.Results: More samples were positive using milk ring test (MRT) (55.6%, 45/81) than using enzyme-linked immunosorbent assay (ELISA) (29.6%, 24/81), with all ELISA positive samples also positive in MRT. The ELISA results were used for risk factor analyses. Seropositive farms were significantly (p = 0.015) larger than seronegative, with an average 7.9 cattle, compared to 4.9. Seropositive farms were more likely to report stillbirth occurring the last year, and a significantly higher proportion of seropositive farms reported retained placenta (odds ratio 5.2).Conclusion: This study showed that Brucella seroprevalence is high among farms in Haryana, and a control program is needed to ensure improved human and animal health, as well as improved livestock productivity.

Comparison of ELISA and Urine Microscopy for Diagnosis ofSchistosoma haematobiumInfection

BackgroundSchistosoma haematobium which causes urogenital schistosomiasis (UGS) is highly prevalent in African countries. Urine microscopy (UM) is the first-line diagnostic method of UGS. Enzyme-linked immunosorbent assay (ELISA) is a common method for screening many parasite infections primarily or alternatively. The present study established an in-house diagnostic system by ELISA and evaluated its diagnostic efficacy in comparison with UM for screening UGS in White Nile State, Republic of Sudan, 2011–2013.MethodsA total of 490 participants were screened by UM or ELISA, and 149 by both. The in-house ELISA system was established employing soluble egg antigen of S. haematobium and the cut-off absorbance was set at 0.270.ResultsOf the 149 subjects, 58 participants (38.9%) were positive by UM, 119 (79.9%) were positive by ELISA and 82 (55.0%) showed consistently positive or negative results by both methods. The diagnostic sensitivity of ELISA was 94.8% and specificity was 29.7% based on UM results. The ELISA positive serum samples also cross-reacted with egg antigens of Schistosoma mansoni and Schistosoma japonicum.ConclusionWe have established in-house ELISA for screening serum immunoglobulin (Ig) G antibodies by employing soluble egg antigen of S. haematobium for diagnosis of UGS with 94.8% sensitivity and 29.7% specificity. The ELISA system can supplement the conventional diagnosis by UM.

Occurrence, seasonal variation and risk assessment of exposure to aflatoxin M1 in Iranian traditional cheeses

A total of 360 traditional cheeses consisted of Lighvan (n = 62), Koozeh (n = 62), Siahmazgi (n = 58), Khiki (n = 58), Talesh (n = 58) and Lactic (n = 62) collected from different parts of Iran were analyzed for aflatoxin M1 (AFM1) using an enzyme linked immunosorbent assay (ELISA). Frequency of AFM1 and its concentration ranges of all the ELISA positive samples were determined by high-performance liquid chromatography with fluorescence detection (HPLC-FD). AFM1 was detected in 60.3%, 75.8%, 72.4%, 43.5%, 38.7% and 35.4% of Siahmazgi, Khiki, Talesh, Lighvan, Koozeh and Lactic cheeses, respectively with concentration ranging from 50.5 to 308.7 ng/kg, respectively. HPLC analyses confirmed the ELISA results although the rates of contaminated cheese samples were lower than that of ELISA. There was significant difference in AFM1 level between various cheese types and samples collected from summer and winter seasons (P < 0.05). By comparing our findings with the EU limit, about 10.5% of cheese samples had exceeding values for the toxin. The results of the present study indicates that there is no health risk in consumption of Iranian traditional cheeses due to the presence of AFM1.

Serotyping of Dengue Virus from Deadly Outbreaks of Pakistan

Dengue virus (DENV) belonging to the family Flaviviridae consists of 4 serotypes (DEN-1, 2, 3 & 4) causing severe illnesses like dengue fever (DF), Dengue Hemorrhagic Fever (DHF) and Dengue Shock Syndrome (DSS). Dengue fever has become an emerging disease in Pakistan during the past 5-7 years with an increase in the number of cases every year since 2006. The current study aimed at identifying the serotypes of dengue virus involved in 2011 outbreak of Pakistan. Samples were collected from all provinces of Pakistan. Patients were confirmed in the hospital by antibody captured Enzyme linked immunosorbent assay (ELISA). These ELISA positive serum samples were inoculated on HeLa cell line to raise the viral titer of dengue virus and then RT-PCR was done for serotyping of dengue virus. Initially D1, D2 primers were tried for serotyping of dengue virus but poor results were obtained. Then serotype specific primers were designed having equal fragment length targeting NS1 gene. This approach was successful in serotyping of dengue virus from Pakistani population. On Real time RT-PCR, Dengue virus (DENV) was detected in 43.33% (n=26) patientsout of 60 analyzed samples. Among 26 positive cases, 76.9% (n=20) were children and 23.1% (n=6) were adultsand 46.1% (n=12) were male and 53.8 % (n=14) were females. ELISA results of26 PCR positive cases show that 53.8% (n=14) were positive for IgM, 15.4 % (n=4) were positive for IgG and 30.8 % (n=8) were positive for both IgG and IgM. In this study DEN-2 was detected in 13.3% (n=8) patients, DEN-3 was detected in 16.67% (n=10) patients and DEN-2 & DEN-3 coinfection was detected in 13.3% (n=8) patients. We did not detected DEN-1 & DEN-4 in 2011 outbreak of Pakistan in 60 analyzed samples. By this study we can conclude that DEN-2 and DEN-3 were the main serotypes involved in 2011 outbreaks in Pakistan.

Aflatoxin M1 in milk and traditional dairy products from west part of Iran: Occurrence and seasonal variation with an emphasis on risk assessment of human exposure

In the present study, a total of 358 samples consisting of raw milk of cow (n = 64), goat (n = 56) and sheep (n = 52); traditional cheese (n = 40), yoghurt (n = 42), Kashk (n = 40), Doogh (n = 44) and Tarkhineh (n = 20) were analyzed for aflatoxin M1 (AFM1) by using an enzyme linked immunosorbent assay (ELISA). Frequency of AFM1 and its concentration ranges in the ELISA positive samples were determined by high performance liquid chromatography with fluorescence detection (HPLC-FD). AFM1 contamination was 84.3%, 44.6% and 65.3% for cow, goat and sheep raw milks, respectively. Moreover, AFM1 was in 65.5%, 23.8%, 14%, 13.6% and 35.0% of cheese, yoghurt, Kashk, Doogh and Tarkhineh samples, respectively. Percentages of cow milk, goat milk, sheep milk and cheese samples exceeding the EU limit were 35.9%, 11.1%, 26.9% and 10%, respectively. HPLC analyses confirmed the ELISA results although the percentages of AFM1 contamination in raw milk and dairy products were lower than that of ELISA. There were significant differences (P < 0.05) between the mean AFM1 contents of raw milk, cheese and yoghurt samples during winter and summer seasons. Our study demonstrated that there is a potential risk for liver cancer due to the consumption of milk and dairy products in Iranian consumers.

Prevalence of Hepatitis G Virus Among Hemodialysis and Kidney Transplant Patients in Khuzestan Province, Iran

Background:Hepatitis G virus (HGV) is a member of Flaviviridae. Prevalence of HGV in healthy people is very low, but this virus is more prevalent in patients with hepatitis. Besides, relative frequency of HGV in patients undergoing hemodialysis, and kidney recipients is very high. The role of HGV in pathogenesis is not clear. Since this virus cannot be cultivated, molecular techniques such as Revers Transcription Polymerase Chain Reaction (RT-PCR) is applied to detect HGV.Objectives:The current study aimed to investigate the prevalence of HGV using determination of E2, viral envelope antigen, antibodies and the RNA by Enzyme Linked Immunosorbent Assay (ELISA) and RT-PCR techniques. The rational of the study was to determine the prevalence of HGV in patients undergoing hemodialysis and kidney transplantation in Khuzestan province, Iran.Patients and Methods:Five hundred and sixteen serum samples of the patients undergoing hemodialysis and kidney transplantation from various cities of Khuzestan province were collected. Anti-hepatitis G E2 antibodies were investigated by ELISA method. RNAs were extracted from serums and Hepatitis G RNA was detected by RT-PCR.Results:Of the 516 samples, 38 (7.36%) specimens were positive for anti-HGV by ELISA. All of these ELISA positive samples were negative for HGV genome by RT-PCR. Of the remaining 478 ELISA negative samples, 16 (3.14%) samples were positive by RT-PCR.Conclusions:Hepatitis G Virus was not prevalent in the patients undergoing hemodialysis and kidney transplantation in Khuzestan province. Although reports indicated high frequency of co-infection of HGV with hepatitis B and C viruses, in the current research, co-infection of HGV with B and C was not considerable. Since different groups and subtypes of HGV are reported, periodic epidemiologic evaluation of HGV and its co-infection with other hepatitis viruses is suggested in other populations such as the patients with thalassemia; however, periodic epidemiologic monitoring of HGV may be helpful to control future potential variations of the virus.

First report of Bovine Viral Diarrhea Virus antigen from pneumonic cattle in Sudan

To explore the expected role of Bovine Viral Diarrhea Virus (BVDV) in pneumonia in cattle, cattle lungs (n=242) showing signs of pneumonia were collected from slaughter houses of three different localities located at Northern, Central and Western Sudan during 2010–2013. The collected samples were tested for the presence of BVDV antigen using Enzyme-Linked Immunosorbent Assay (ELISA), and Fluorescent Antibody Test (FAT). Twenty six (10.7%) out of 242 samples were found to be positive for BVDV. Positive results were seen in all the three studied areas, with the highest prevalence (16.7%; n=4/24) at Gezira State in Central Sudan. BVDV genome could be detected in all ELISA positive samples. The results indicated the existence of BVDV infection in cattle in different areas in Sudan, and its possible association with respiratory infections in cattle. Analysis using BLAST indicated that the sequence was identical to the previously reported BVDV-1 (GenBank accession AF220247.1.); nucleotide A was found in our study at position 9 of our sequence, whereas T was present instead in the reference virus. This is the first report of detecting BVDV antigen, genome, and its sequence analysis collected from cattle lungs in Sudan.