大熊猫轮状病毒RT-PCR检测方法的建立

Establishment of RT-PCR for detection of giant panda rotavirus

根据GenBank中登录的大熊猫轮状病毒(RV)VP7基因序列,设计了1对特异性引物,建立了快速检测大熊猫RV的RT-PCR方法。用该方法对MA-104细胞增殖的大熊猫RV进行RT-PCR扩增,结果得到与试验设计相符的342bp的特异性扩增条带,而对传染性胃肠炎病毒、大肠杆菌和沙门氏菌的扩增结果为阴性。测序比对结果证实该体系检测结果准确;该方法最低可检测出约1pg的病毒核酸;重复性试验结果显示,其检测重复性好,提示该RT-PCR方法可用于大熊猫RV的临床快速检测。

Based on the published sequence of the giant panda rotavirus strain CH-1 VP7 gene in GenBank,a pair of specific primers was designed,and a rapid RT-PCR method was established for detecting rotavirus in giant panda.The specific band of 342bp was amplified from the culture of MA-104,but no speci-fic band was amplified from transmissible gastroenteritis virus,Escherichia coli and Salmonella.Sequence analysis indicated that this method was accurate.As little as 1pg RNA of giant panda rotavirus could be detected,and repeatability was good.In conclusion,the established RT-PCR method could be used for the detection of rotavirus in giant panda.