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目的建立套式引物聚合酶链反应（ＰＣＲ），用于检测滤纸干血斑中的人免疫缺陷病毒１型（ＨＩＶ１）前病毒ｐｏｌ基因ＤＮＡ片段。方法采集ＨＩＶ１感染者的全血约５０μｌ滴在经ＥＤＴＡ－蛋白酶Ｋ预处理并干燥的滤纸片上，室温下干燥，将滤纸片密封于塑料袋中，在室温及４℃下保存１～６４周后，分别将滤纸片置０５ｍｌ试管中直接进行ＨＩＶ１前病毒ｐｏｌ基因的外侧引物ＰＣＲ检测，然后进行内侧引物的ＰＣＲ检测。结果经ＥＤＴＡ－蛋白酶Ｋ预处理的滤纸片在４℃下保存４０周、在室温下保存２４周仍可检出ＨＩＶ１前病毒目的基因。根据ＰＣＲ产物的琼脂糖凝胶电泳溴化乙锭染色带形并参比实验设立的标准对照可直接判断结果。结论该方法具有快速、特异、敏感的特点，敏感性可以达到检出１０个靶ＤＮＡ分子。样品采集后可通过邮件传递至中心实验室，特别适合于ＨＩＶ１感染的确证及筛检
Objective To establish a nested primer polymerase chain reaction (PCR) for the detection of DNA fragment of pol gene of human immunodeficiency virus type 1 (HIV1) in filter paper dried blood spots. Methods Fifty μl of whole blood from HIV-1 infected patients were collected on filter paper pretreated with EDTA-proteinase K and dried. After drying at room temperature, the filter paper was sealed in a plastic bag and stored at room temperature and 4 ° C for 1 to 64 weeks , Respectively, the filter paper set 0 5ml test tube directly to the outside of the pol gene of HIV1 virus PCR primer PCR, and then carry out PCR primer detection. Results The filter paper pretreated with EDTA-proteinase K was stored at 4 ℃ for 40 weeks, and the target gene of HIV1 virus was still detected after being stored at room temperature for 24 weeks. According to the PCR product of agarose gel electrophoresis ethidium bromide staining and reference set up a standard test can be directly judged by the results. Conclusion The method is rapid, specific and sensitive, and its sensitivity can detect up to 10 target DNA molecules. After the sample is collected, it can be transmitted to the central laboratory by mail, which is especially suitable for confirmation and screening of HIV1 infection