[Study on Rotavirus detection by nucleic acid sequence based amplification].
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.
Kou
Xiao-Xi
XX
Guangdong Instisute of Microbiology, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangzhou 510070, China. kou_er@163.com
Wu
Qing-Ping
QP
Fan
Hong-Ying
HY
D017931:DNA Primers / Q000235:genetics; D005243:Feces / Q000821:virology; D005759:Gastroenteritis / Q000821:virology; D006801:Humans; D012401:Rotavirus / Q000235:genetics / Q000302:isolation & purification*; D012400:Rotavirus Infections / Q000821:virology; D021161:Self-Sustained Sequence Replication / Q000379:methods*; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000235:genetics
D017931:DNA Primers; D014764:Viral Proteins