[Study on Rotavirus detection by nucleic acid sequence based amplification].

Xiao-Xi Kou; Qing-Ping Wu; Hong-Ying Fan

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PubMed · 16736602

[Study on Rotavirus detection by nucleic acid sequence based amplification].

Xiao-Xi Kou·Qing-Ping Wu·Hong-Ying Fan

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Abstract

Rotavirus is one of the major cause of the viral gastroenteritis throughout the world. A nucleic acid sequence-based amplification(NASBA) technique for the detection of rotavirus in faecal specimens was developed and compared to the RT-PCR technique. Primers were designed according to the high conserved region of VP7 gene. Amplicons were detected by denaturating agarose gel, and then demonstrated by Northern hybridization with digoxigenin-labeled oligonucleotide probe. The anticipative specific amplification product is 392bp,and no nonspecific products appear even the concentration of nontarget nucleic acid as high as 1 microg/microL. A detection limit of 50 pg target RNA/mL is obtained when the optimal amplification time of 3h used.The NASBA assay will be a favourable alternative to RT-PCR for the investigation of rotavirus outbreaks as a routine diagnostic test in the near future because it is shown to be highly sensitive, specific and do not require specialized equipment.

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DNA Primers Feces Gastroenteritis Humans Rotavirus Rotavirus Infections Self-Sustained Sequence Replication Sensitivity and Specificity Viral Proteins Evaluation Study Journal Article Research Support, Non-U.S. Gov't

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Xiao-Xi Kou, Qing-Ping Wu, Hong-Ying Fan. 2006. [Study on Rotavirus detection by nucleic acid sequence based amplification].. https://pubmed.ncbi.nlm.nih.gov/16736602/

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