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目的验证商品化寨卡病毒核酸测定试剂盒,建立赣州市寨卡病毒的检测方法,为寨卡病毒疫情的判定及制定防控措施提供依据。方法用商品化寨卡病毒核酸测定试剂盒,对中国CDC检测定性的6份标本进行回顾性检测,用荧光定量PCR方法检测病毒核酸拷贝数浓度。对寨卡阳性标准品(1×10~7copies/ml)和以10倍梯度稀释为10~(-1)至10~(-7)倍共8个浓度梯度分别检测,验证试剂盒敏感性,对Ct值和拷贝浓度进行拟合获得标准曲线。对麻疹、风疹、登革热、肠道EV71型和肠道CA16型病毒进行核酸定性检测,验证试剂盒特异性。结果敏感性验证实验最低检测限约为1×10~3copies/ml;对麻疹等5种病毒检测定性为阴性;6份标本中病毒拷贝数为:1.9×10~4copies/ml、<0、2.7×10~5copies/ml、<0、<0、<0;且阴阳性对照和内部对照均成立。结论商品化试剂盒对本次寨卡病毒阳性血清和尿液标本可以定性检出,符合预期,对其他5种病毒无交叉反应。有一定的敏感性和特异性。试剂盒可以作为本市寨卡病毒的初步筛查检测,可用于寨卡病毒疫情的初步判定及防控。
Objective To validate the commercialized Zika virus nucleic acid assay kit and establish the detection method of Zika virus in Ganzhou city so as to provide the basis for the judgment of Zika virus epidemic situation and the formulation of prevention and control measures. Methods Commercially available Zika virus nucleic acid assay kit was used to detect 6 qualitative samples of CDC in China. The copy number of the virus nucleic acid was detected by fluorescence quantitative PCR. The sensitivity of the kit was tested against 8 sets of concentrations ranging from 1 × 10 ~ 7 copies / ml and 10 ~ (-1) to 10 ~ (-7) The Ct value and copy concentration were fitted to obtain a standard curve. Qualitative detection of measles, rubella, dengue, enteric EV71 and enteric CA16 viruses was performed to verify the specificity of the kit. Results The minimum detectable limit of sensitivity test was 1 × 10 ~ 3 copies / ml. The detection of 5 kinds of viruses such as measles was qualitatively negative. The virus copy number in 6 samples was 1.9 × 10 ~ 4 copies / ml, <0,2.7 × 10 ~ 5 copies / ml, <0, <0, <0; and negative control and internal control were established. Conclusion The commercial kit can qualitatively detect the positive serum and urine samples of Zika virus, which is in line with expectation and has no cross-reaction with the other five viruses. Have a certain sensitivity and specificity. The kit can be used as a preliminary screening test of Zika virus in the city and can be used for the preliminary determination and prevention and control of the epidemic situation of Zika virus.