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Rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification.

ABSTRACT: BACKGROUND: PKV is a new emerging pathogen detected in diarrhea pigs. At present, no more detection methods were reported except RT-PCR method. this study was to develop a fast diagnostic method based on the LAMP reaction for rapid detection of PKV nucleic acid in fecal samples. FINDINGS: Two pairs of primers were designed to amplify the conservative 3D gene of PKV genome. The PKV RT-LAMP method possessed well specificity and had 100 times higher sensitivity than common reverse transcription PCR (RT-PCR), which could detect up to 10 RNA copies of the target gene. CONCLUSIONS: The results showed that the optimal reaction condition for RT-LAMP was achieved at 64°C for 50 min. Furthermore, the RT-LAMP procedure does not demand special equipment and is time-saving.

SUBMITTER: Li C

PROVIDER: S-EPMC4026823 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification.

Li Changlong C Chen Jianfei J Shi Hongyan H Zhang Xin X Shi Da D Han Xiao X Chi Yanbin Y Feng Li L

Virology journal 20140423

<h4>Background</h4>PKV is a new emerging pathogen detected in diarrhea pigs. At present, no more detection methods were reported except RT-PCR method. this study was to develop a fast diagnostic method based on the LAMP reaction for rapid detection of PKV nucleic acid in fecal samples.<h4>Findings</h4>Two pairs of primers were designed to amplify the conservative 3D gene of PKV genome. The PKV RT-LAMP method possessed well specificity and had 100 times higher sensitivity than common reverse tran ...[more]

PMID: 24755372

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