PMID: 18724714Aug 30, 2008Paper

Development and application of real-time quantitative RT- PCR assay for the detection of hepatitis E virus

Sheng wu gong cheng xue bao = Chinese journal of biotechnology
Caixia QiaoXiangying Duan

Abstract

Hepatitis E virus (HEV) sequences including four major genotypes representative strains available in GenBank were aligned with the DNAMAN software. The highly conserved internal region of ORF2 was then subjected to design primers and a probe. Furthermore, a 0.3 kb fragment of HEV ORF2 containing the amplification region was transcribed in vitro to synthese cRNA standard and a universal real-time TaqMan PCR assay was optimized and developed to detect and quantify main genotypes RNA of HEV. The specificity and reliability of the real-time RT-PCR was confirmed by testing genotype I HEV, genotype IV HEV and clinical samples. The detection limit of real-time RT-PCR was found 2.0 x 10(1) copies per reaction using in vitro transcribed cRNA. Compared with nested RT-PCR in diagnosis of HEV, the real-time RT-PCR developed was 10 to 100-fold more sensitive than the nested RT-PCR. The detection results from 54 clinical specimens indicated real-time RT-PCR was a rapid, sensitive and reproducible diagnostic method for HEV. This assay will be useful as an early and rapid diagnostic assay for HEV.

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.