Enteric non‐A, non‐B hepatitis: Epidemics, animal transmission, and hepatitis E virus detection by the polymerase chain reaction

Jameel, Shahid; Durgapal, Hemlata; Habibullah, C. M.; Khuroo, Mohammed Sultan; Panda, Subrat Kumar

doi:10.1002/jmv.1890370405

Cited by 101 publications

(68 citation statements)

References 17 publications

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“…IgM antibody to ORF-1, ORF-2, and ORF-3 of HEV and HEV RNA were tested by methods developed at our institute. 13,14 Anti-hepatitis C antibody was tested by using a third-generation, commercial enzyme-linked immunosorbent assay (Xcyton, Bangalore, India) method. Hepatitis C virus RNA was tested by reverse transcription nested polymerase chain reaction, using primers from the 5Ј nontranslated region, if appropriate by methods already described.…”

Section: Methodsmentioning

confidence: 99%

A 20-year single-center experience with acute liver failure during pregnancy: Is the prognosis really worse?

et al. 2008

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Pregnant patients with acute liver failure (ALF) are believed to have a worse outcome than nonpregnant women and men with ALF. However objective data supporting this supposition are scant. Therefore, the current study compared the outcome, complications, and causes of ALF among pregnant women and girls with age-matched nonpregnant women and girls and men and boys with ALF. One thousand fifteen consecutive ALF patients in the reproductive age group, admitted at the All India Institute of Medical Sciences, New Delhi, from January 1986 to December 2006, were included in the study. A total of 249 (38.5%) women were pregnant. They were compared with 341 nonpregnant women and girls and 425 men and boys, aged 15 to 45 years. The mortality rate of pregnant women and girls (53.8%) was similar to age-matched nonpregnant women and girls (57.2%), and men and boys (57.9%); P ‫؍‬ 0.572.The clinical and biochemical features, disease severity, and complications were also similar in the three groups. A significantly higher proportion of ALF was attributable to hepatitis E virus (HEV) among women and girls who were pregnant (59.4%), as compared with both nonpregnant women and girls (30.4%), and men and boys (23.1%); P < 0.001. However, the outcome of HEV-related ALF was independent of the sex and pregnancy status of the patients (P ‫؍‬ 0.103). Mortality in HEV-ALF and non-HEV-ALF patients in pregnant women and girls was 51% (74/145) and 54.7% (52/ 95)(P > 0.1), respectively. The outcome of pregnant ALF patients was also unrelated to the trimester of pregnancy. The mortality of non-HEV-related ALF among the pregnant women and girls (54.7%), age-matched nonpregnant women and girls (61.7%), and men and boys (62.8%) were also similar (P > 0.1). Conclusion: The mortality of pregnant patients with ALF is similar to that of nonpregnant women and girls and men and boys and is independent of the cause or trimester. Pregnancy per se should not be regarded as a poor prognostic factor for a patient with ALF. (HEPATOLOGY 2008;48:1577-1585

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Section: Methodsmentioning

confidence: 99%

A 20-year single-center experience with acute liver failure during pregnancy: Is the prognosis really worse?

et al. 2008

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“…The HEV clone ORF1-pSGI was digested with EcoRI and XbaI to release an insert ranging from nt 1 to nt 4449, whereas the insert ranging from nt 4449 to nt 7195 was released from the nt 4438 to nt 7195 pBluescript SK(ϩ) clone by digestion with restriction enzymes XbaI and XhoI. These inserts were cloned into a pSGI vector (13) digested with restriction enzymes EcoRI and XhoI in a three-way ligation. A schematic representation of the reconstruction is given in Fig.…”

Section: Methodsmentioning

confidence: 99%

The In Vitro-Synthesized RNA from a cDNA Clone of Hepatitis E Virus Is Infectious

Panda

Ansari

Durgapal

et al. 2000

J Virol

124

117

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Hepatitis E virus (HEV) is an important etiological agent of epidemic and sporadic hepatitis, which is endemic to the Indian subcontinent and prevalent in most of the developing parts of the world. The infection is often associated with acute liver failure and high mortality, particularly in pregnant women. In order to develop methods of intervention, it is essential to understand the biology of the virus. This is particularly important as no reliable in vitro culture system is available. We have constructed a cDNA clone encompassing the complete HEV genome from independently characterized subgenomic fragments of an Indian epidemic isolate. Transfection studies were carried out with HepG2 cells using in vitro-transcribed RNA from this full-length HEV cDNA clone. The presence of negative-sense RNA, indicative of viral replication, was demonstrated in the transfected cells by strand-specific reverse transcription-PCR and slot blot hybridization. The viral proteins pORF2 and pORF3 and processed components of the pORF1 polyprotein (putative methyltransferase, helicase, and RNA-dependent RNA polymerase) were identified in the transfected cells by metabolic pulse-labeling with Hepatitis E virus (HEV) is established as an etiological agent of the epidemic and sporadic forms of waterborne hepatitis (6, 25). The first well-characterized HEV epidemic was reported in Delhi, India, in 1955 (35). Several other epidemics have since been described in developing countries on nearly every continent (19,21). HEV has a positive-strand polyadenylated RNA genome ϳ7.2 kb in length (27) containing three open reading frames (ORFs). Nonstructural ORF1 (5Ј end) codes for a polyprotein of 1,693 amino acids (pORF1) and contain domains homologous to a viral methyltransferase, a papainlike cysteine protease, an RNA helicase, and an RNAdependent RNA polymerase (RdRp). The second ORF (3Ј end) codes for the major viral capsid protein of 660 amino acid (pORF2), while the third and smallest ORF (ORF3) codes for a 123-amino-acid-long polypeptide (pORF3) whose function is unknown (32). Apart from its coding region, the viral genome has 27-and 68-nucleotide (nt)-long noncoding regions at its 5Ј and 3Ј ends, respectively (32). The genome sequences of HEV have been reported from different geographical isolates and show a high degree of homology at both the nucleotide and amino acid levels (3,4,12,32,33). Expression of structural proteins pORF2 and pORF3 in prokaryotic and eukaryotic systems has been reported by different investigators (7,11,14,23,29). Earlier, we have expressed and characterized pORF2 and pORF3 in animal cells. pORF2 is an 88-kDa glycoprotein which is expressed intracellularly, as well as on the cell surface (14, 37). The ORF3 protein (pORF3) is a 13.5-kDa phosphoprotein which is phosphorylated by the cellular mitogen-activated protein kinase and associates with the cytoskeleton (36). We have also recently expressed the ORF1 polyprotein in both prokaryotic and eukaryotic systems (2).In the absence of a reliable in vitro culture system...

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“…[12][13][14] The primer sequences are given in Table 1. The PCR for HBV was seminested while that of HCV and HEV was nested.…”

Section: Primersmentioning

confidence: 99%

Multiplex Reverse Transcriptase-PCR for Simultaneous Detection of Hepatitis B, C, and E Virus

Garg

Kumar

Asim

et al. 2016

Journal of Clinical and Experimental Hepatology

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Introduction: The hepatitis B virus (HBV), HCV, and HEV may occur as singly or concurrently in patients of different kind of liver disease. The rapid, reliable, and cost-effective screening of these pathogens is required for the large epidemiological studies. Therefore, a study has been planned to develop a multiplex Reverse Transcriptase-PCR assay which can be used for the screening of maximum number of pathogens at a time. Methodology: To develop multiplex Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) assay for simultaneous detection of HBV, HCV, and HEV; the serum samples of 54 patients who were positive either singly or in co-infection with for HBV, HCV, and HEV serologically were screened by uniplex PCR/RT-PCR followed by multiplex RT-PCR for HBV, HCV, and HEV using specific primers. These primers can detect most genotypes of these viruses. Multiplex RT-PCR was done in one tube for the identification of viral DNA/RNA using a mixture of three pairs of specific primers for hepatitis B, C, and E viruses. Representative positive samples of these viruses by uniplex/multiplex RT-PCR were also confirmed by sequencing followed by alignment with reference strains sequence. Results: The specificity of multiplex PCR was 100% with high sensitivity 89%, 87%, and 74% for HBV, HCV, and HEV respectively. The sensitivity and specificity of RT-multiplex PCR demonstrated a good correlation with that of uniplex PCR. Conclusion: The study suggests that multiplex RT-PCR can serve as a simple and reliable assay for rapid, sensitive, and cost-effective method for simultaneous detection of super-infections with HEV particularly in Asian countries as a cause of decompensation of chronic liver disease. ( J CLIN EXP HEPATOL 2016;6:33-39)

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Enteric non‐A, non‐B hepatitis: Epidemics, animal transmission, and hepatitis E virus detection by the polymerase chain reaction

Cited by 101 publications

References 17 publications

A 20-year single-center experience with acute liver failure during pregnancy: Is the prognosis really worse?

A 20-year single-center experience with acute liver failure during pregnancy: Is the prognosis really worse?

The In Vitro-Synthesized RNA from a cDNA Clone of Hepatitis E Virus Is Infectious

Multiplex Reverse Transcriptase-PCR for Simultaneous Detection of Hepatitis B, C, and E Virus

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