Comparison of the effectiveness of different nucleic acid detection kits in detecting avian influenza virus H5 subtypes in environmental samples
Objective To compare the detection efficiencies of real-time reverse transcription-polymerase chain reaction(Real-time RT-PCR)kits from three manufacturers for detecting avian influenza virus H5 subtype on environmental specimens.Methods Kits from three companies were used to detected H5 subtype avian influenza virus of the samples from live poultry market.Statistical methods were used to compare the difference in the detection rate of different kits.The PCR products were subjected to TA cloning and sequencing.Results Among the kits from three companies,the positive rate of H5 by using kits from company C was higher than that of company A(18.3％vs.8.4％),the difference was statistically significant(x2=5.440,P=0.039).TA cloning and sequencing showed that the sequence of PCR products of kit A shows highest similarity with sequence KU042758.1(100.00％).Sequence of PCR products of kit C shows highest similarity with sequence CY091635.1(97.92％).Conclusion Kit C had a higher detection rate of H5 in this batch of clinical samples than kit A,which might be caused by the inconsistency of the target sequence of different kits and the matching degree of the kit primer to the nucleic sample acid.
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