[Study on rotavirus detection with single-tube seminested RT-PCR method in shellfish].
Being a foodborne virus, Rotavirus was often carried by shellfish. At the present, RT-PCR was the most effective method of Rotavirus detection in shellfish, but its sensitivity was low because of low levels of virus contamination and PCR inhibitors in shellfish. So contaminated shellfish experimentally in laboratory, and imitated the natural environment to concentrate Rotavirus, then detected by the developed single-tube seminested RT-PCR. Compared with ELISA and the conventional RT-PCR, the detection sensitivity is improved by 1000 times and 10 times respectively. In addition, the outer and inner contamination is reduced dramatically and the detection time is decreased from 6h to 4.5h. When the whole shellfish and only the digestive tissues of shellfish serve as samples, the detection ratio and sensitivity are more higher when sample is the latter.
Guangdong Institute of Microbiology, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangzhou 510070, China. kou-er@163.com
Xiao-xia
XX
Kou
Qing-ping
QP
Wu
Jv-mei
JM
Zhang
Hong-ying
HY
Fan
D000818:Animals; D004279:DNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012401:Rotavirus / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D012758:Shellfish / Q000821:virology*
D004279:DNA, Viral