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作者用计算机辅助分析和比较16种HIV-1和HIV-2序列并设计了一对用于检测HIV-l和HIV-2的通用PCR引物PG03和PG04.它们在ARB-2病毒序列上分别位于457~478和766~798位核苷酸。两引物在ARV-2基因上指导合成一个长342bp的DNA片段。用pARV-2/7A和含有gag基因的亚克隆质粒pJG423作模板,均可扩增出特异性靶基因序列,而用不含gag基因的亚克隆质粒pJE332,pJP163和pJP032作模板则无扩增产物。将pJG423进行稀释,结果该PCR法可检出少至数个拷贝的质粒DNA,表明此法敏感性极高。作者用PCR法还直接从HIV阳性血清中检测出HIV基因序列,而正常对照血清却不能扩增出PCR产物。
The authors used computer-aided analysis and comparison of 16 HIV-1 and HIV-2 sequences and designed a pair of universal PCR primers PG03 and PG04 for the detection of HIV-1 and HIV-2. They are located at nucleotides 457-47 and 766-798 of the ARB-2 viral sequences, respectively. The two primers guide the synthesis of a 342 bp DNA fragment on the ARV-2 gene. Specific target gene sequences were amplified using pARV-2 / 7A and subclone plasmid pJG423 containing gag gene as template, but no amplification was carried out with subclone plasmids pJE332, pJP163 and pJP032 without gag gene as templates product. The pJG423 was diluted, the results of this PCR method can detect as few copies of the plasmid DNA, indicating that this method is extremely sensitive. The authors also detected HIV gene sequences directly from HIV-positive sera by PCR, whereas normal control sera did not amplify PCR products.