摘要
目的针对WHO-NEQAS提供的20份已知疟原虫血样在不同实验室应用不同PCR诊断方法的检测准确率数据进行统计分析和比较,并运用巢氏PCR对上述样本进行检测准确率的效果评价。方法采用配对T检验的统计学方法比较不同实验室对已知性质和数量的疟原虫样本,在巢氏PCR、多重PCR和荧光定量PCR 3种方法检测准确率之间的差异性;运用巢氏PCR对20份样本进行PCR扩增和琼脂糖凝胶电泳。结果在不考虑疟原虫种类时,巢氏PCR和多重PCR相对于荧光定量PCR的检测准确率较高且均存在显著差异(P=0.0033,P=0.0456);在原虫密度小于10^(3)虫/ml血时,巢氏PCR和多重PCR相较于荧光定量PCR在检测准确率上均存在显著差异(P=0.0014,P=0.0125);在原虫密度为10^(4)~10^(5)虫/ml血以及大于10^(6)虫/ml血时,3种方法之间并无显著差异(P>0.05)。结论巢氏PCR用于鉴别部分疟原虫种诊断,特别是在原虫密度较低时,仍具有较高的检测准确率和灵敏度,结果更为可靠。
Objective To analyze and compare the detection rate of 20 Plasmodium blood samples provided by WHO-NEQAS using different PCR diagnostic methods in different laboratories,and evaluate the effect of nested-PCR on the detection coincidence rate.Methods The statistical method of paired t-test was used to compare the differences in the detection accuracy of nested PCR,multiplex PCR,and fluorescence quantitative PCR in different laboratories;20 samples were amplified and conducted agarose gel electrophoresis by using nested-PCR.Results When the species of Plasmodium were not considered,the detection rates of nested-PCR and multiplex PCR were higher than that of real-time PCR(P=0.0033,P=0.0456).When the parasitemia was less than 10^(3)parasites/ml blood,there was a significant difference in the detection rate between nested-PCR and multiplex PCR compared with real-time PCR(P=0.0014,P=0.0125).There was no significant difference between the three methods when the parasitemia was 10^(4)-10^(5)or more than 10^(6)parasites/ml blood(P>0.05).Conclusion Nested-PCR is a more reliable detection method for its high detection rate and sensitivity to identify certain Plasmodium species for diagnosis,especially when the parasitemia is low.
作者
张逸龙
叶润
张冬梅
ZHANG Yi-long;YE Run;ZHANG Dong-mei(Department of Tropical Diseases,Faculty of Naval Medicine,Naval Medical University,Shanghai 200433,China)
出处
《中华卫生杀虫药械》
CAS
2022年第4期345-351,共7页
Chinese Journal of Hygienic Insecticides and Equipments
基金
国家自然科学基金青年基金(编号:31601002)
上海市2019年度“科技创新行动计划”(编号:19JC1416500)
关键词
巢氏PCR
疟疾检测
诊断方法
效果评价
nested-PCR
malaria detection
diagnostic methods
effect evaluation
