五种布鲁氏菌核酸检测试剂盒检测性能的评价北大核心CSCD

目的比较5种布鲁氏菌核酸实时荧光PCR检测试剂盒的一致性和检出能力,为临床实验室选择检测方法和布鲁氏菌的诊断提供参考依据。方法选用经病原学检测确定为布鲁氏菌阳性的血液样本38份,健康人的血液样本24份,潘氏变形杆菌、溶藻弧菌、河弧菌、铜绿假单胞菌、肺炎克雷伯菌DNA各1份,使用5种试剂盒(编号A-E)分别进行核酸检测,比较5种试剂盒临床样本检测的一致性;选择1份阳性样本核酸用无RNA酶水梯度稀释得到5个浓度(浓度1:4453.13 fg/μL,浓度2:1113.28 fg/μL,浓度3:278.32 fg/μL,浓度4:69.58 fg/μL,浓度5:17.40 fg/μL),每个浓度使用5种试剂盒(编号A-E)分别进行3次检测,比较5种试剂盒的阳性检出率及批内重复性。结果5种试剂盒检测67份DNA样品的符合率稍有不同,试剂盒ABDE的符合率均为100%,试剂盒C的符合率为98.51%。批内重复性显示5种试剂盒在浓度1、浓度2、浓度3水平重复检测DNA的Ct值变异系数均<5%;在浓度1与浓度4梯度区间,试剂盒的阳性检出能力比较显示试剂盒A、B、D较高,为11/12,试剂盒C和E较低,为8/12。结论5种试剂盒的真实性和可靠性较好,灵敏度和符合率稍有差别,特异度均为100%;重复性较好,检测性能良好。部分试剂盒对弱阳性样本的检出能力不强,该类样本可使用多种试剂盒复核,以保障结果的准确性。

Evaluation of the detection capability of five Brucella nucleic acid detection kits

This study compared the consistency and detection capability of five Brucella detection kits (real-time PCR method) to provide a reference for the selection of detection methods in clinical laboratories. To compare the consistency of the five kits, we selected 38 blood samples confirmed to be Brucella positive by etiological tests, 24 blood samples from healthy people, and the DNA of five other bacteria for nucleic acid detection. To compare the positive detection rate and repeatability of the five kits, we selected one positive nucleic acid sample, which was diluted to five concentrations in RNase free water. Each concentration was tested three times with the five kits (A-E). The consistency rate of 67 DNA samples detected by the five kits slightly differed. Kit C’s consistency rate was 98.51%, whereas the coincidence rates of the other four kits were 100%. Intra-batch repeatability analysis indicated that the coefficients of variation of DNA Ct values repeatedly detected by the five kits were less than 5% at concentrations 1-4. In the gradient range of concentrations 1-4, the kits’ positive detection capability was higher for kits A, B and D (11/12), and lower for kits C and E (8/12). These five kits’ validity and reliability were good, their sensitivity and consistency rates slightly differed, and their specificity was 100%. The repeatability and detection capability were good. However, some kits’ detection capabilities were insufficient to detect weakly positive samples. Such samples could be rechecked with a variety of kits to ensure the accuracy of the results.