Easy-to-use rapid gene amplification method for direct detection of RNA and DNA viruses in sera and feces from various animals.

Abstract:

:The development of rapid and simple gene amplification tests is required for detection of pathogens to prevent transmission of infectious diseases between animals or from animals to humans. An easy-to-use rapid gene amplification method that can directly detect RNA and DNA viruses in clinical samples was developed. This method is based on combining loop-mediated isothermal amplification (LAMP) or reverse transcription-LAMP (RT-LAMP) and RNA GEM Tissue, a thermophilic enzyme that extracts nucleic acid by quickly digesting proteins and ribonucleases. The authors named these methods GEM LAMP and GEM RT-LAMP. These methods were able to detect viral DNA and RNA within 70 min in a single tube using only a water bath. The detection capacities were 10-100-fold more sensitive than those of previously established LAMP and RT-LAMP methods. The GEM LAMP and GEM RT-LAMP methods were used to detect macroscopically the presence of DNA and RNA viruses in sera or fecal samples from cattle, pigs, horses, dolphins, penguins, and sea lions using SYBR green I. The GEM LAMP and GEM RT-LAMP methods thus have considerable versatility as tools for detecting pathogens and are applicable to basic human and veterinary medicine, environmental hygiene, and point-of-care-testing.

journal_name

J Virol Methods

authors

Segawa T,Kobayashi Y,Sase Y,Itou T,Suzuki M,Endoh T,Nakanishi T,Sakai T

doi

10.1016/j.jviromet.2014.01.019

subject

Has Abstract

pub_date

2014-06-01 00:00:00

pages

31-7

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(14)00029-9

journal_volume

201

pub_type

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