Risk of hepatitis-E virus infections among blood donors in a regional blood transfusion centre in western India.

BACKGROUND: Hepatitis-E virus (HEV) is an emerging infectious threat to blood safety. The enormity of the transmission of HEV and its clinical consequence are issues currently under debate. This study aimed to evaluate the prevalence of HEV-RNA in blood donors in western India. MATERIALS AND METHODS: We screened 13 050 blood donors for HEV using HEV-RNA screening of 10 mini-pools using RealStar HEV RT-PCR Kit (95% limit of detection (LOD): 4.7 IU/ml). Furthermore, all HEV-RNA-positive donors were investigated for the presence of IgM/IgG antibody along with liver function tests. RESULTS: Of the 13 050 blood donations, 7 (0.53%) were found to be HEV-RNA positive, and the prevalence of HEV nucleic acid testing yield cases among blood donors was 1 in 1864. All seven HEV-RNA-positive samples were tested with anti-HEV IgM and anti-HEV IgG antibodies; this resulted in two (28.5%) positive anti-HEV IgM and two (28.5%) positive anti-HEV IgG antibodies. Hepatic activity was measured, with two of seven HEV-RNA-positive donors demonstrating abnormal serum glutamic oxaloacetic transaminase (SGOT) andserum glutamic pyruvic transaminase (SGPT). Two HEV-RNA-positive blood donors who had abnormal SGOT and SGPT were found to have a high HEV viral load. Furthermore, we were able to follow up two HEV-RNA donors, and both were HEV-RNA positive and had anti-HEV IgM and anti-HEV IgG antibodies; moreover, their liver function tests were also abnormal. One of the HEV-RNA donors with high viral load did show hepatitis-E-like virus on electron microscopy. CONCLUSION: Our studies indicate that there is a significant risk of blood-borne transmission of HEV. This finding may help to provide a direction towards the safety of blood transfusions in clinical settings in countries like India, which fall under the endemic category for HEV infection.

A multiplex ARMS PCR approach to detection of common ß-globin gene mutations.

BACKGROUND: ß-thalassaemia is a group of inherited single-gene disorders worldwide. Each ethnic population has its own common mutations. Heterogeneity of ß-thalassaemia mutations in multi-ethnic population of Surat, makes molecular diagnosis expensive and time consuming. METHODS: Specific primers were used to differentiate four common mutations, IVS I-5 (G→C), Codon 41/42 (- TCTT), 619-bp deletion and FS 8/9 (+G), by a simple PCR involving a multiplex amplification refractory mutation system. RESULTS: Several high prevalence ß-Thalassemia trait groups constituted by Muslims, Patels, Sindhis, ModhBanias, and Mahayavanshi. Four most common mutations detected in them are IVS I-5 (G→C), Codon 41/42 (- TCTT), 619-bp deletion and FS 8/9 (+G). We identified each of these ß-thalassemia mutations in multiplexed ARMS from positive control samples. Our multiplex-ARMS-PCR system was first standardized on positive DNA samples with above known four most common ß-thalassemia mutations, and these positive samples had been diagnosed with ß-thalassemia and also all these samples belonged to Surat ethnic groups. The system was subsequently tested on 110 blood samples from different ethnic backgrounds with unknown ß-thalassemia mutations which were in all specimens. CONCLUSION: The ARMS multiplex system was found reliable, cost effective, fast and most applicable for mutation screening of Thalassemia in Surat populations.

NAT positivity in seronegative voluntary blood donors from western India.

BACKGROUND AND OBJECTIVES: Prevalence and composition of Hepatitis B, Hepatitis C and HIV-1, NAT positive but seronegative voluntary blood donors from western part of India is yet to be documented. MATERIAL AND METHODS: Over last 2 1/2 years all the seronegative voluntary blood donors were tested using 10 minipools on a semiautomated NAT testing platform. The positively tested donors were followed up for at least five months for development of seropositivity. RESULTS: 79532 seronegative donations were tested by 10 minipool (MP) NAT leading to 51 positive sample (44 Hep B, 5 HIV 1 and Hep C positive). All the HIV and Hep C NAT positive donors eventually developed seropositivity and out of 44 Hep B NAT positive donors, 31 developed seropositivity within six months of follow up, following counseling of the donors. This data translate into NAT yield of 1:1559 donors for all virus taken together. NAT yield for Hep B 1:1807 donors were much higher than HIV 1 in 1:15906 and HCV yield of 1:39761. Semiautomated minipool NAT testing system was found to be cost effective way for improving blood safety. INTERPRETATION AND CONCLUSION: Seronegative NAT yield in voluntary blood donors are quiet high in western part of India and in line with rest of the country is mainly due to Hepatitis B infection. Implementation of strict donor screening, Hep B vaccination of the population and sample mutation of NAT testing should be under taken on war footing.

Development of an animal model of Helicobacter pylori (Indian strain) infection.

To develop an animal model for Indian strain Helicobacter pylori (H. pylori) infection. This model will allow one to study many facts of H. pylori infection in a more controlled manner. Mongolian gerbils were orogastric inoculated with two different Indian strains of H. pylori at different time points. Animals were sacrificed and looked for the presence of infection up to 52 weeks post-inoculation using a variety of techniques. Simultaneously, serums from these animals were also tested for antibody, and changes in the histopathology of stomach on H&E (hematoxylin and eosin) stains were also noted. Experimental sets of Mongolian gerbils were orally fed two strains of H. pylori obtained from human case by culture of different cagA and vacA strains three times daily on days 0, 2, and 4. H. pylori ATCC26695 strain was used for antisera preparation; three animals from each group were sacrificed at different time periods 2, 4, 8, 12, 26, 38 and 52 weeks after infection along with one control animal. Infections with H. pylori were confirmed in all the animals from 4 weeks onwards up to 52 weeks with histopathological changes in conformity with H. pylori gastritis. Wild Mongolian gerbils can be infected with Indian strains of H. pylori, and the infection persists at least 1 year. However, intensity of gastritis was milder than that seen in human case.