猕猴逆转录病毒多重套式PCR检测方法的建立

Establishment of Multiple Nested-PCR Detection for Rhesus Monkey's Retroviruses

分别针对编码STLV-1和SRV/D-1两种逆转录病毒膜蛋白的env基因进行引物设计,通过优化、调整PCR条件,建立一种能同时检测猕猴STLV-1和SRV/D-1两种逆转录病毒的多重PCR方法,用于猕猴种群逆转录病毒的常规监测。结果显示多重套式PCR产物片段大小与预期结果一致,进一步测序证实为目的产物,说明建立的多重套式PCR方法能同时检测出猕猴体内可能存在的STLV-1和SRV/D-1两种逆转录病毒。这种方法具有灵敏度高、特异性强、省时、试剂用量少和检测费用低等优点,因此可以作为一种新方法用于猕猴种群逆转录病毒的定性监测。

In order to establish a multiple nested-PCR method that can be used for monitoring SRV/D-1 and STLV-1 simultaneously in captive rhesus monkey colony, two pairs of specific primers respectively for SRV/D-1 and STLV-1 according to retroviruses＇ conserved env gene region were designed. By adjusting and optimizing the PCR condition, the two retroviruses can be detected in the same PCR system at one time. The gene segments of the multiple nested-PCR products were 476 bp and 422 bp, and the gene sequencing were STLV-1 and SRV/D-1 respectively. The study showed that SRV/D-1 and STLV-1 could be detected simultaneously using our multiple nested-PCR, which had a lower cost, high sensitivity and specificity for monitoring retroviruses in rhesus monkey colony.