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目的比较3种禽流感病毒荧光定量反转录-聚合酶链式反应(RT-PCR)检测试剂盒。方法采集禽流感外环境标本40份,通过标本检测、阳性标本倍比稀释检测以及扩增产物交叉扩增试验,比较3种试剂盒的检测结果、灵敏度和稳定性。结果 A、B和C试剂盒,每份样品的检测用时分别为94、56和106 min;阳性检出率分别70.0%、45.0%和45.0%,差异有统计学意义(χ~2=9.750,P=0.045)。C试剂盒灵敏度最高,其次为B和A试剂盒。3种试剂盒核酸检测结果的变异系数均小于5%。扩增产物交叉扩增试验结果显示,其中任一种试剂盒的初次扩增产物都可被另外两种试剂盒再次扩增。结论 3种试剂盒的稳定性均较好,但灵敏度、成本、操作等存在一定的差异,建议实验人员结合实验室条件及试验目的合理选择试剂。
Objective To compare three fluorescent quantitative reverse transcription - polymerase chain reaction (RT-PCR) detection kits. Methods Forty samples of peripheral blood samples of bird flu were collected. The samples were tested, the ratio of dilution to positive samples and the cross-amplification test of amplification products were compared. The test results, sensitivity and stability of the three kinds of kits were compared. Results The detection time of each sample was 94, 56 and 106 min for the A, B and C samples, respectively. The positive detection rates were 70.0%, 45.0% and 45.0% respectively, with significant difference (χ ~ 2 = 9.750, P = 0.045). C kit has the highest sensitivity, followed by B and A kit. The coefficients of variation of nucleic acid test results of the three kits were less than 5%. Amplification products The results of the cross-amplification test showed that the primary amplification products of any of the kits can be re-amplified by the other two kits. Conclusion The stability of the three kinds of kits is good, but the sensitivity, cost, operation and so there is a certain difference, it is recommended that the experimenter reasonable selection of reagents based on laboratory conditions and test purposes.