Tags

Type your tag names separated by a space and hit enter

A broadly reactive one-step real-time RT-PCR assay for rapid and sensitive detection of hepatitis E virus.
J Virol Methods. 2006 Jan; 131(1):65-71.JV

Abstract

Hepatitis E virus (HEV) is transmitted by the fecal-oral route and causes sporadic and epidemic forms of acute hepatitis. Large waterborne HEV epidemics have been documented exclusively in developing countries. At least four major genotypes of HEV have been reported worldwide: genotype 1 (found primarily in Asian countries), genotype 2 (isolated from a single outbreak in Mexico), genotype 3 (identified in swine and humans in the United States and many other countries), and genotype 4 (identified in humans, swine and other animals in Asia). To better detect and quantitate different HEV strains that may be present in clinical and environmental samples, we developed a rapid and sensitive real-time RT-PCR assay for the detection of HEV RNA. Primers and probes for the real-time RT-PCR were selected based on the multiple sequence alignments of 27 sequences of the ORF3 region. Thirteen HEV isolates representing genotypes 1-4 were used to standardize the real-time RT-PCR assay. The TaqMan assay detected as few as four genome equivalent (GE) copies of HEV plasmid DNA and detected as low as 0.12 50% pig infectious dose (PID50) of swine HEV. Different concentrations of swine HEV (120-1.2PID50) spiked into a surface water concentrate were detected in the real-time RT-PCR assay. This is the first reporting of a broadly reactive TaqMan RT-PCR assay for the detection of HEV in clinical and environmental samples.

Authors+Show Affiliations

Centers for Disease Control and Prevention, National Center for Infectious Diseases, 4770 Buford Highway, Mailstop F-36, and Emory University, Department of Global Health, Rollins School of Public Health, Atlanta, GA 30341, USA. JIN2@cdc.govNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

16125257

Citation

Jothikumar, Narayanan, et al. "A Broadly Reactive One-step Real-time RT-PCR Assay for Rapid and Sensitive Detection of Hepatitis E Virus." Journal of Virological Methods, vol. 131, no. 1, 2006, pp. 65-71.
Jothikumar N, Cromeans TL, Robertson BH, et al. A broadly reactive one-step real-time RT-PCR assay for rapid and sensitive detection of hepatitis E virus. J Virol Methods. 2006;131(1):65-71.
Jothikumar, N., Cromeans, T. L., Robertson, B. H., Meng, X. J., & Hill, V. R. (2006). A broadly reactive one-step real-time RT-PCR assay for rapid and sensitive detection of hepatitis E virus. Journal of Virological Methods, 131(1), 65-71.
Jothikumar N, et al. A Broadly Reactive One-step Real-time RT-PCR Assay for Rapid and Sensitive Detection of Hepatitis E Virus. J Virol Methods. 2006;131(1):65-71. PubMed PMID: 16125257.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A broadly reactive one-step real-time RT-PCR assay for rapid and sensitive detection of hepatitis E virus. AU - Jothikumar,Narayanan, AU - Cromeans,Theresa L, AU - Robertson,Betty H, AU - Meng,X J, AU - Hill,Vincent R, Y1 - 2005/08/24/ PY - 2005/05/19/received PY - 2005/07/11/revised PY - 2005/07/14/accepted PY - 2005/8/30/pubmed PY - 2006/4/14/medline PY - 2005/8/30/entrez SP - 65 EP - 71 JF - Journal of virological methods JO - J Virol Methods VL - 131 IS - 1 N2 - Hepatitis E virus (HEV) is transmitted by the fecal-oral route and causes sporadic and epidemic forms of acute hepatitis. Large waterborne HEV epidemics have been documented exclusively in developing countries. At least four major genotypes of HEV have been reported worldwide: genotype 1 (found primarily in Asian countries), genotype 2 (isolated from a single outbreak in Mexico), genotype 3 (identified in swine and humans in the United States and many other countries), and genotype 4 (identified in humans, swine and other animals in Asia). To better detect and quantitate different HEV strains that may be present in clinical and environmental samples, we developed a rapid and sensitive real-time RT-PCR assay for the detection of HEV RNA. Primers and probes for the real-time RT-PCR were selected based on the multiple sequence alignments of 27 sequences of the ORF3 region. Thirteen HEV isolates representing genotypes 1-4 were used to standardize the real-time RT-PCR assay. The TaqMan assay detected as few as four genome equivalent (GE) copies of HEV plasmid DNA and detected as low as 0.12 50% pig infectious dose (PID50) of swine HEV. Different concentrations of swine HEV (120-1.2PID50) spiked into a surface water concentrate were detected in the real-time RT-PCR assay. This is the first reporting of a broadly reactive TaqMan RT-PCR assay for the detection of HEV in clinical and environmental samples. SN - 0166-0934 UR - https://www.unboundmedicine.com/medline/citation/16125257/A_broadly_reactive_one_step_real_time_RT_PCR_assay_for_rapid_and_sensitive_detection_of_hepatitis_E_virus_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0166-0934(05)00241-7 DB - PRIME DP - Unbound Medicine ER -