Generation ELISA Research Articles

Prevalence and Trends of Hepatitis B, Hepatitis C and Human Immunodeficiency Viruses among Blood Donors in Islamabad, Pakistan 2005-2013

Background: Pakistan is a developing country of 190 million people with increased burden of infectious diseases. Hepatitis B virus (HBV), Hepatitis C virus (HCV) and Human Immunodeficiency virus (HIV) are the most important agents responsible for transfusion transmitted viral infections. Blood donors are considered as the healthiest population of a community and screening of HBV, HCV & HIV in blood donors will reflect the true prevalence of these infections in a population. Material and Methods: The study was conducted from July 2005 to July 2013 at Pakistan Institute of Medical Sciences (PIMS) hospital, Islamabad, Pakistan. 160,376 blood donors from the age 18 years to 60 years were enrolled in this study. Study was approved by the ethical review committee of PIMS hospital. All the samples were screened for Hepatitis B virus surface Antigen (HBsAg), anti-HCV and anti-HIV by fourth generation ELISA. Statistical analyses were carried out by using SPS software version 17. Results: Among 160,376 donors, 157,920 (98.47%) were replacement donors. The overall seroprevalence of (HBVsAg), anti-HCV and anti-HIV among blood donors were 2.35%, 3.26% & 0.017% respectively. The prevalence of HBV and HCV co-infection was 0.084%. Conclusion: The prevalence of Hepatitis B, Hepatitis C and Human Immunodeficiency viruses are high in blood donors and should not be acceptable. There is a need to educate masses to increase the number of voluntary donors to meet the country’s requirement of blood donation

Sero-prevalence of HTLV 1/2 and HCV in paired mother and children from Malawi as well as systematic review and meta-analysis of HTLV-1/2 and HCV of data available on women and children living in Africa

HTLV prevalence is poorly mapped in several areas, in particular in Africa. Global prevalence is commonly cited as being 10-20 million but more recently this has been contested as an overestimation. Additional epidemiological studies would further our understanding of the HTLV burden. Similarly, HCV is endemic in certain locations (Egypt) whilst data for other areas of Africa are virtually absent. We investigated HTLV/HCV prevalence in serum samples from 418 paired mother and child samples of healthy mothers of children attending Malawian hospitals. Latest generation ELISAs were used; consistently ELISA reactive mothers’ sera were further tested by immunoblot and only deemed sero-positive if they were reactive in this assay. Serum from the children of ELISA positive/borderline mothers were subsequently screened. Sero-prevalence in the healthy women was as follows, HTLV-1: 0.72% (n=3), HTLV-2: 1.9% (n=8), HTLV indeterminate: 3.8% (n=16) and HCV: 0.48% (n=2). DNA extracted from whole blood could only resolve one of the high percentage of intermediate positive results, most likely because of a low proviral load. One child in each category: HTLV-1, HTLV-2, dual-infected and HTLV indeterminate was identified indicating low (11.54%) mother to child transmission (MTCT) whereas one of the two HCV positive mothers had a HCV positive child, suggestive of high HCV MTCT transmission rate. We conducted a systematic review to compare this original data with previous reports: Malawi had similar, low, levels of HTLV-1 and HCV but surprisingly high HTLV-2. Prevalence maps for HTLV and HCV for this demographic in the African continent and the meta-analysis will be presented.

Risk factors for hepatitis C virus sero-positivity among haemodialysis patients receiving care at kidney centre in a tertiary health facility in Maiduguri, Nigeria.

Hepatitis C virus (HCV) is an important health care problem in haemodialysis. Hepatitis C virus is both a cause and complication of kidney diseases. Yet there are limited information on antibody against HCV in patients on haemodialysis. The purpose of this study was to determine the prevalence of anti-HCV and the risk factors associated with HCV infection in a cohort of 100 participants on haemodialysis. They were consecutively recruited into the study, anti-HCV testing was made by the 3rd-generation ELISA System (C-100, C-33c, C-22). The prevalence of HCV antibody was 15%, risk factors associated with HCV antibody were history of blood transfusion and duration of session of haemodialysis; the risk increased with increased with the number of blood transfusion and seasons of haemodialysis. The observed high prevalence of HCV antibody among patients on haemodialysis reflect the quality of healthcare services and the standards of infection control practices in our haemodialysis units. Routine screening for HCV should be done before blood transfusion using third generation ELISA assays with high sensitivity and specificity. Safety measures should be taken in our haemodialysis units to prevent cross infection among patients and staffs. These safety measures include; discarding syringes, needles, gloves, bloodlines and dialysers after single use, and the use of sterile dressings on each patient visit.

Prevalence of hepatitis B e antigen among human immunodeficiency virus and hepatitis B virus co-infected patients in Jos, Nigeria

Human immunodeficiency virus (HIV) negatively impacts the natural history of hepatitis B virus (HBV) infection, including replication. We determined the prevalence of HBeAg in HIV/HBV co-infected patients compared to HBV mono-infected controls and further investigated the relationship between HBeAg seropositivity and the degree of HIV-induced immunosuppression in co-infected patients. The study design was cross-sectional. One hundred HBsAg-positive HIV-infected adults and 100 age and sex matched HBsAg-positive HIV negative controls were consecutively recruited between May and November 2010. Relevant demographic and HBV-related information was obtained. HBeAg was assayed by semi-quantitative third generation ELISA. The HIV/HBV co-infected patients also had CD4+ cell and HIV viral load quantification measured using flow cytometry and polymerase chain reaction techniques respectively. In each group, the mean age was 34 ± 8 years and the majority (61%) was female. The prevalence of HBeAg was significantly higher among co-infected patients (n = 28; 28%) than in the controls (n = 15; 15%; p = 0.03). HBeAg seropositivity was independently associated with age < 40 years (AOR = 2.83, 95% = CI 1.29-6.17) and HIV seropositivity (AOR = 2.44, 95% C.I = 1.17-5.07). The prevalence of HBeAg was significantly higher in co-infected patients with CD4 cell count < 200 cell/µL (41.3%) compared to those with 200-499 cell/µL (18.6%) and≥500 cell/µL (9.1%), p = 0.006. HIV/HBV co-infected patients have a significantly higher prevalence of HBeAg than HBV mono-infected individuals. HBV-infected patients should be routinely assessed for HBeAg, especially if they are co-infected with HIV.

Risk factors for Hepatitis C virus antibody seropositivity among children with sickle cell anaemia in Ilorin, Nigeria

Background : Hepatitis C is an infectious disease of the liver caused by the hepatitis C virus (HCV) resulting to a chronic hepatitis. Chronic HCV infection constitutes a serious public health challenge in Nigeria where donor blood is not routinely screened for HCV. Patients with sickle cell anaemia (SCA) are considered a subset of the population at higher risk of acquiring the virus, due to their frequent needs for transfusion of blood and its products. Other risk factors like scarification markings, tattooing, and circumcision also predispose children to acquiring this infection. However, the magnitude of HCV infection has not been adequately measured in our general population and specific data on HCV in SCA patients are scanty, hence a prospective case controlled study to determine the risk factors that predispose to the acquisition of hepatitis C Virus infection. Objective : To determine the risk factors for Hepatitis C Virus Antibody Seropositivity among transfused children with SCA in Ilorin. Subjects and Method : Eighty two transfused SCA children aged 6 months to 14 years were recruited consecutively from February 2008 to January 2009 while eighty four non transfused SCA children of the same age range recruited over the same period served as controls. Hepatitis C virus Antibody screening was done using a second generation ELISA method. Information on the study population were collected by use of a pretested questionnaire by the investigator. Results : There was a positive correlation between numbers of units of blood transfused and seropositivity. Those who had three or more units of blood had a prevalence rate of more than 50%. There was a strong correlation between seropositivity and scarification marks in both subjects and controls (p=0.001 and 0.02 respectively). Other plausible risk factors for hepatitis C infection tested in this study included circumcision and sharing of clippers which showed no statistically significant difference. No cases of tattooing, drug abuse, needle sharing or sexual activities were seen in this study. Conclusion : Transfused SCA patients belong to a high risk group for hepatitis C virus infection compared to the nontransfused population. The risk of acquisition increases with higher number of transfusions and scarifications marks. Key words : Hepatitis C virus, Sickle cell anaemia, Risk factors, Blood transfusion.

HCV Envelope protein 2 sequence comparison of Pakistani isolate and In-silico prediction of conserved epitopes for vaccine development

BackgroundHCV is causing hundreds of cases yearly in Pakistan and has become a threat for Pakistani population. HCV E2 protein is a transmembrane protein involved in viral attachment and thus can serve as an important target for vaccine development but because of its variability, vaccine development against it has become a challenge. Therefore, this study was designed to isolate the HCV E2 gene from Pakistani HCV infected patients of 3a genotype, to perform In-silico analysis of HCV E2 isolated in Pakistan and to analyze HCV E2 protein sequence in comparison with other E2 proteins belonging to 3a and 1a genotypes to find potential conserved B-cells and T-cell epitopes that can be important in designing novel inhibitory compounds and peptide vaccine against genotype 3a and 1a.Patients and methodsPatients were selected on the basis of elevated serum ALT and AST levels at least for six months, histological examination, and detection of serum HCV RNA anti-HCV antibodies (3rd generation ELISA). RNA isolation, cDNA synthesis, amplification, cloning and sequencing was performed from 4 patient’s serum samples in order to get the HCV E2 sequence. HCV E2 protein of Pakistani origin was analyzed using various bioinformatics tools including sequence and structure tools.ResultsHCV E1 protein modeling was performed with I-TASSER online server and quality of the model was assessed with ramchandran plot and Z-score. A total of 3 B-cell and 3 T-cell epitopes were found to be highly conserved among HCV 3a and 1a genotype.ConclusionThe present study revealed potential conserved B-cell and T-cell epitopes of the HCV E2 protein along with 3D protein modeling. These conserved B-cell and T-cell epitopes can be helpful in developing effective vaccines against HCV and thus limiting threats of HCV infection in Pakistan.

Expression of Matrix Metalloproteinase 1( MMP1 ) in Hepatocellular Carcinoma ( HCC ) : Immunohistochemical and Biochemical Studies

Background: The present study aimed to evaluate the expression of Matrix Metalloproteinase-1 (MMP1) in HepatoCellular Carcinoma (HCC) by using the Immunohistochemical technique, which allows us to integrate the biological aspects of this enzymatic expression in the morphological context of HCCs. Material and Methods: The study was performed on 70 subjects from out and in patients of Tropical medicine Department, Thiodor Billhars Institute during the period from January 2011 until June 2012. The present study included 60 patients with chronic hepatitis C who had undergone liver biopsy. They consisted of 42 men and 28 women with ages ranging from 36 to 66 years. The diagnosis of chronic hepatitis C was made on the basis of positivity for anti-HCV (by the second generation ELISA), and confirmed by HCV-RNA reverse transcription polymerase chain reaction (RT-PCR). Patients were divided into four groups: Group I: included 10 normal persons with no history of liver disease with normal liver enzymes and free ultrasonographic finding as normal control. It included 6 males, 4 females, with ages ranging from 34 to 48 years. Group II: included 20 HCV infected patients without cirrhotic changes. It included 11 males, 9 females, with ages ranging from 39 to 53 years. Group III: included 20 HCV infected patients with liver cirrhosis, 12 males, 8 females, with ages ranging from 48-63 years. Group IV: included 20 HCV infected patients with HCC, 16 males, 4 females, with ages ranging from 53-64 years. Results: Blood Picture, (Hb, WBCs, RBCs, Plts, PC and ESR). Liver Function Test (ALT, AST, ALB, GGT, ALP, T. BIL and D. BIL). Matrix Metalloprotenase 1(MMP1) Measurements: Serum MMP1. Histopathological investigation Including histopathological changes in the liver tissue. Conclusion: our results suggest that MMP-1 is overexpressed in a large proportion of patients with HCC and the high expression level of protein correlated with the disease progression and poor clinical outcome in HCC. Furthermore, MMP-1 high expression proved to be a risk factor for tumor recurrence and independent molecular marker of prognosis in HCC and may become a novel target in the strategies for the prediction of tumor progression and prognosis of this disease.

The ETR (End Treatment Response) of Chinese Brand Human recombinant interferon alpha 2a plus Ribavirin in chronic hepatitis C patients selected from Combined Military Hospitals in Pakistan

This Study was carried out to determine the End of Treatment Response (ETR) of a Chinese Source Human Recombinant Interferon Alpha 2a and Ribavirin in chronic hepatitis C affected patients, selected from different combined military hospitals in Pakistan. In this study hepatitis affected noncirrhotic patients from different combined military hospitals in Pakistan were selected from September 2010 to April 2011. Sixty seven patients were selected based on the inclusion exclusion criteria. Fifty seven patients completed the 24 weeks therapy for Hepatitis C. The Patients (n=57) had persistently raised serum aminotransferase (ALT), positive HCV antibodies by 3rd generation ELISA, positive HCV RNA by polymerase chain reaction. Study group patients underwent 24 weeks therapy with a Chinese source Interferon and Ribavirin and subsequently followed up for the End of Treatment Response by the assay of HCV RNA by polymerase chain reaction at 24 weeks. Patients of chronic hepatitis C (44 males and 13 females) had age range 20-56 years. After 24 weeks of therapy with a Chinese source Interferon and Ribavirin, 73.7% patients showed a favourable ETR manifested by Negative HCV RNA polymerase chain reaction.Treatment with a Chinese source Interferon and Ribavirin combination therapy for 24 weeks produces a favourable End of Treatment Response in patients of chronic hepatitis C from different military hospitals in Pakistan.

Prevalence of HBs Ag among blood donors in Transfusion Center, Abidjan (Ivory Coast)

Aim: To study the prevalence of HBsAg among persons without risk factors for hepatitis B. Patients and Methods: Cross-sectional study (28 January to 31 December 2010) about 4310 new volunteers blood donors consecutively recruited. Among all, a risk factor for hepatitis B was investigated (standardized questionnaire and clinical examination). Blood of all blood donors without risk factors of hepatitis B (n = 3823) were systematically collected for HBsAg, hepatitis C antibody, syphilis serology and human immunodeficiency virus serology. The blood was analyzed with a third generation ELISA (Abbott Murex). Results: HBsAg was positive in 179 persons (4.68% [95% CI 4% - 5%]). It was associated with hepatitis C antibodies (0.28% (n = 11)), syphilis antibodies (0.68% (n = 26)) or anti-human immunodeficiency virus antibodies (0.2% (n = 8)). In multivariate analysis, age 30 to 39 years (1.545 [95% CI 1.074 to 2.222]), male sex (2.426 [95% CI 1.550 to 3.799]) and positive syphilis serology (14.344 [95% CI 5.161 to 39.865]) were associated with presence of HBsAg. Conclusion: The prevalence of HBsAg among adult without risk factors for hepatitis B was not negligible. Also, it is essential to maintain routine screening before any vaccination against hepatitis B in adult. Our study is not for an immunization against hepatitis B only targeting risk populations in adult.

Hepatitis C virus co-infection among people living with HIV/AIDS in a Nigerian Teaching Hospital

AimThis study is the first attempt conducted to determine the co-infection rates of Hepatitis C virus among people living with HIV/AIDS (PLWHA) in our hospital. This is expected to lead to improvement in treatment protocols. BackgroundCo-infection of Hepatitis C virus infection and HIV (Human Immune Deficiency Virus) infection is not to be unexpected as they share routes of infection. The detection of such patients is particularly important as the treatment may involve life threatening drug interactions. This could result in shorter life expectancy of the patients. Materials and MethodsThis is a prospective study. A total of 280 subjects living with HIV/AIDS were recruited into this study. Sera of the participants were subjected to anti-HCV antibody (IgG) screening using third generation ELISA kit from DIA.PRO, Italy. The prevalence of HCV was determined and bar charts and tables were utilised to depict the prevalence rates of various groups. ResultsSixty five (23.2%) of the participants were found to be co-infected. No association was established between age, sex, partners status, previous transfusion, and number of partners. ConclusionThe prevalence of 23.2% for HCV among PLWHA reported for this community is alarming. More focus should be directed on management of HCV component of coinfection than it is done now. Further studies are recommended employing molecular techniques to determine subtypes of HCV that may be predominant and why this figure of co-infection is so high in the study location.

Hepatitis C Viral Prevalence and Seroconversion in Moroccan Hemodialysis Units: Eight Year Follow Up

Introduction: The prevalence of Hepatitis C Virus (HCV) in chronic hemodialysis patients is eight times higher than among the general population. It is estimated between 3 and 65.8% in different studies and countries and is a contributor to mortality in this population. The aim of this study is to define the factors influencing prevalence and seroconversion of HCV in an ambispective study including 15 centers and 163 hemodialysis patients. Methods: Multicenter ambispective Study including 167 patients treated in 15 dialysis centers in 4 cities and over a period of 8 years. In 2002, all patients underwent determination of liver enzymes, a serological survey and qualitative research of HCV RNA by polymerase chain reaction followed by a genotyping for patients confirmed positive by the method LIPA. Serological status and movement of patients between centers was followed for 8 years. In 2010, the survivors benefit serology of viral hepatitis B and C (4th generation ELISA) with research of viral RNA by PCR. Results: The mean age of patients at baseline was 51 years of which 11% were diabetic with a mean of 55 months on hemodialysis. The prevalence of HCV was 33.4% with significantly higher ALT in positive patients and genotype 1b was most frequent (54%). Statistical analysis found that the factors of prevalence are: time on hemodialysis, number of units visited (2.2 vs. 3, p<0.001), number of red blood cells transfused (1.5 vs. 2.5, p=0.009) and area of the unit. After 8 years, overall survival was 58.2% with a prevalence of HCV at 26.2%. During follow-up, 7 seroconversion were objectified and the only factor found is the number of centers visited. Conclusions: These data suggest that nosocomial transmission plays an important role hence the importance of strict implementation of the recommendations of prevention against the transmission of HCV in particular the training of personnel and control the transfer of patients between centers.

Hepatitis C virus antibodies among transfused children with sickle cell anaemia at University of Ilorin Teaching Hospital

Background: Hepatitis C is an infectious disease of the liver caused by the Hepatitis C virus (HCV) resulting to a chronic Hepatitis. Chronic HCV infection constitutes a serious health challenge in places where prevalence is substantial. In Nigeria, there is a high risk because donor blood is not routinely screened for HCV. Patients with sickle cell anaemia (SCA) are considered a subset of the population at higher risk of acquiring the virus, due to their frequent needs for transfusion of blood and its products. However, the magnitude of HCV infection has not been adequately measured in our general population and specific data on HCV in SCA patients are scanty, hence a prospective case controlled study to determine the prevalence of HCV antibodies in transfused SCA patients attending the sickle cell anaemia clinic in the University of Ilorin Teaching Hospital (UITH), Ilorin was taken. Objective: To determine the prevalence of Hepatitis C virus antibodies among transfused children with SCA in Ilorin. Subjects and Method: Eighty two transfused SCA children aged 6 months to 14 years were recruited consecutively from February 2008 to January 2009 while eighty four non transfused SCA children of the same age range recruited over the same period served as controls. Hepatitis C virus antibody screening was done using a second generation ELISA method. Results: The overall prevalence of HCV antibody was 3.0%, while it was 3.7% and 2.4% in the transfused and non transfused SCA patients respectively (χ2 = 0.23, p = 0.68). The patients were also comparable across the social class when subcategorized into high and low social class (χ2 = 0.37, p = 1.00 (subjects), χ2 = 0.42, p = 1.00 (controls). Conclusion: The prevalence of Hepatitis C virus anti- bodies in transfused SCA patients is low. The difference in prevalence between transfused and nontransfused SCA patient was not statistically significant. This was cautiously interpreted due to the hospital based premise of the work. Therefore, Hepatitis C virus antibody acquisition might be from sources other than transfusion of unscreened blood.

Study the prevalence of torque teno (TT) virus infection in patients with hematological malignancies

Torque Teno (TT) virus infection may increase the risk of different types of leukemia. In this study the infective markers of TT virus, hepatitis B virus (HBV), and hepatitis C virus (HCV) were evaluated in patients with different types of leukemia in comparison with controls. In this cross sectional study Ethylenediaminetetraacetic acid (EDTA)-treated blood samples were collected from 163 patients with leukemia and 100 control between years: 2007-2011. Molecular prevalence of TT virus infection was evaluated in studied populations using in-house semi-nested polymerase chain reaction (PCR) protocol. HBsAg and HCVAb were analyzed in studied groups by third generation ELISA kits according to manufacturer’s instructions. The genomic DNA of TT virus was found in 29 of 163 (17.8%) of leukemia patients and 2 of 100 (2%) of controls. HBsAg was diagnosed in 21 of 147 (14.3%) patients with leukemia and in 2 of 100 (2%) controls. HCVAb was diagnosed in 4 of 147 (2.7%) patients with leukemia and in none of controls. The co-infection of TT virus with HBV and with HCV was found in 4 of 147 (2.7%) and in 1 of 147 (0.7%) of patients with leukemia, respectively. Identification of high prevalence of TT virus infection in patients with leukemia compared with controls emphasize on possible association between TT virus infections with leukemia. Therefore evaluation of the prevalence of TT viral infection in larger groups of patients with long lasting follow up is suggested. Key words: Torque Teno (TT) virus, hepatitis B virus (HBV), hepatitis C virus (HCV), leukemia, hematological malignancies.

Methods for Identification of CA125 from Ovarian Cancer Ascites by High Resolution Mass Spectrometry

CA125 is the most widely used tumour marker in ovarian cancer with unsatisfactory sensitivity and specificity especially at early stage. It is quantified by antibody-based immunoassays; however different molecular weight isoforms have been described in the literature which have never been validated by mass spectrometry, potentially affecting the diagnostic accuracy and clinical reliability of the test. In this study, CA125 was detected by Western blot and its identity confirmed by mass spectrometry. Two-dimensional (2D) gel electrophoresis in combination with mass spectrometry revealed that positive Western blot signals up to 500 kDa are most likely false-positive interactions of M11-like and OC125-like antibodies. Fibronectin, identified as one of these false-positive interaction partners, increased the reading for CA125 in a first generation ELISA significantly (p = 0.02). The existence of low-molecular weight isoforms of CA125 is therefore questionable and is most likely reflecting cross-reactivity of the antibodies with other proteins. This would explain the conflicting reports on the molecular structure of CA125 and also the inconsistency of CA125 levels by different ELISAs. Our results are also the first steps towards a mass spectrometric assay for CA125 quantification, which would improve sensitivity and reliability.

The comparison of the performance between third generation ELISA and fourth generation ELISA on acute and early HIV infection

Objective To evaluate the performance of the third generation ELISA and the fourth generation ELISA for HIV-1 diagnosis assays on acute and early HIV-1 infected samples. Methods Sixty-seven acute/early HIV-1 infected samples were collected from the follow-up gays with seroconversion in Shen Yang city and from clinical patients in the First Affiliated Hospital of China Medical University with incomplete HIV-1 specific bands in western blot between 2008 and 2010.Third generation ELISA, fourth generation ELISA, western blot and HIV-1 viral load detecting were used for detecting these samples.The sensitivity, consistency were compared between third generation ELISA and fourth generation ELISA to detect the seroconversion samples and the window periods were abserved.Chi square test was used for statistical analysis. Results In the 67 acute/early HIV-1 infected samples, 56 were HIV positive and 11 were HIV negative by the third generation ELISA.The sensitivity of the third generation ELISA was 83.6% (95%CI:72.5%-91.5%); 63 were HIV positive，1 was at gray zone and 3 were HIV negative by the fourth generation ELISA.The sensitivity of the fourth generation ELISA was 94.0% (95%CI:85.4%-98.3%),higher than the third generation ELISA（χ2=16.1，P<0.01）.The consistency of the third generation ELISA and the fourth generation ELISA was 86.6%(95%CI：76.0%-93.7%).The earliest third generation ELISA positive sample was the sample collected 16 days after HIV infection and the earliest fourth generation ELISA positive sample was the sample collected 9 days after HIV infection.There was significantly different on the window periods between the third generation ELISA and the fourth generation ELISA. Conclusion The fourth generation ELISA had a higher sensitivity and shorter window period on acute/early HIV infected samples than the third generation ELISA, which is more suitable for the HIV early infection screening on high risk populations. （Chin J Lab Med，2012,35：538-543） Key words: HIV infections; HIV-1; Enzyme-linked immunosorbent assay

HEPATITIS C;

Background: Liver biopsy the gold standard for the histological assessment in chronic hepatitis C is an invasive procedure withcertain limitations. Objective: It was to evaluate the relation of serum ALT levels with the histological changes in healthy blood donors screenedpositive for HCV. Study Design: This cross sectional comparative study Setting: It was conducted at department of medicine CombinedMilitary Hospital Lahore, from January to August 2004 Method: 76 apparently healthy blood donor soldiers age 20 – 49 years positive for HCVantibodies by third generation ELISA test, evaluated by medical history, physical examination, serum ALT levels , PCR test for HCV RNA andultrasound abdomen were enrolled. Liver biopsy was done through percutaneous route and histology was graded according to Knodell scoringsystem. Results: Serum ALT levels showed a skewed distribution with mean of 130.9+96.9 IU/L and median of 8.5IU/L. Eleven patients hadnormal ALT (&lt;40 IU/L).On histological analysis of liver biopsy specimen 17 had Knodell score ≤3 (mild inflammation),38 had score between 4and 7 (moderate inflammation)and 21 had &gt; 8. Patients with score &gt; 8 were older than the former groups.The difference was significant(p=0.047). Relation of ALT levels with stage of fibrosis and grade of inflammation was estimated using ANOVA it showed no statisticalsignificance. Patients with stage 1 fibrosis had highest mean serum ALT levels (p=0.108). Conclusions: No association found between theALT level &amp; liver biopsy histological score. However patients with normal ALT levels usually had mild necroinflammatory changes.

Improving blood safety using fourth generation HIV ELISA as the screening tool in blood banks – an Indian experience

Results The HIV seroprevalence was estimated to be 1.37/1000 donations with 3 generation and 3.62/1000 donations with 4 generation ELISA (p>0.05). Of the 17 samples which were 4 generation ELISA positive and WB positive, 11 were positive with 3 generation ELISA. Fourth generation ELISA tested 19 additional samples positive and 7 samples as possibly positive which were tested negative with 3 generation ELISA, giving an additional yield of 26 window period units per 10200 donations i.e. 2.5/1000 donations. Of these, 6 were WB positive giving a yield of 0.58 window period units per 1000 donations. Since 2-3 components are prepared from each blood unit, the yield can be increased to 12-18/10200 donations. With an annual donation of nearly 46000 blood units, this will be 54-81 positive samples per 10200 donations.

The study of hepatitis B surface antigen and anti-HCV in HIV infected patients

Methods The study group included 106 HIV positive patients with history of heterosexual contact and the control group included 30 asymptomatic healthy individuals. The sera of patients were tested for HBsAg and antiHCV using third generation ELISA kits. The samples positive for first kit were retested for confirmation of results. The sera of control group were simultaneously screened for HIV, HBsAg, and anti-HCV by ELISA method.

Abstract 3205: Isolating RNA, DNA and protein from the same sample allows correlation of multiple biomarkers

Abstract DNA, RNA or proteins are routinely analyzed as markers to define the type and state of tumorigenicity. Each of these analytes provide a unique set of information in order to detect differences such as chromosomal mutations, imbalances in gene expression, and changes in protein signaling networks. Limited samples, such as biopsies and small tumors, are a common sample type, however, due to their small size the analysis of more than one of these analytes from the same sample can be challenging. Taking more than one sample (biopsy), or splitting a sample in order to isolate one of these anlaytes from each section raises questions regarding sample heterogeneity. This is particularly important where one section of a tumor may be different from another, and can even provide misleading information regarding the correlation of DNA, RNA or protein markers. Another issue raised with splitting samples is that two of the analytes are lost when the third is isolated. This can lead to the loss of important information, and the low yield can limit the number and methods used for analysis. Being able to analyze DNA, RNA and proteins from the same sample is particularly crucial during biomarker discovery and correlation. Here we describe a new, quick and easy to use method to isolate DNA, RNA and proteins from small sized tumor and normal adjacent tissue and demonstrate that not only is the entire sample maintained giving us more sample for analysis (no analyte is lost when isolating the other), but there is no concern regarding heterogeneity. Using these samples we demonstrate the correlation of DNA, mRNA, miRNA and protein using Next Generation Sequencing, qPCR, Western and ELISA and associate these with tumorigenicity. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3205. doi:1538-7445.AM2012-3205

Hepatitis C virus infection in hemodialysis patients in Maracaibo, Venezuela

Over a two year period, the incidence of hepatitis C virus (HCV) infection was evaluated in 29 hemodialysis patients, aged between 15 and 75 years (mean ± SD: 45 ± 39.5 years), from the University Hospital Hemodyalisis Unit, Maracaibo, Zulia State, Venezuela. Anti-HCV antibodies were determined using a fourth generation ELISA (Innotest HCV Ab IV) kit and positive blood samples were tested using a recombinant assay kit (Inno-LIA HCV Ab III), both kits from Innogenetics N.V., Belgium. The findings indicate a lack of HCV seroconversion in the hemodialysis patients over the study period, confirmed by the recombinant assay. Risk factors for HCV infection were 0.3270 (95% confidence interval: 0.01323-8.080) in patients undergoing hemodialysis. The findings suggest a lack of significant sources for HCV infection due to the preventive measures to avoid its transmission in the hemodialysis unit.