万孚疟原虫检测试剂盒检测卵形疟原虫效果评价及影响因素分析  

Evaluation of Wondfo Rapid Diagnostic Kit for detecting Plasmodium ovale and analysis of influencing factors

作　　者：唐凤[1] 唐建霞 陆凤[1] 徐岁[1] 顾亚萍[1] 仝德胜[1] 朱国鼎[1] 华海涌[1] 周华云[1] 曹俊[1] 

机构地区：[1]江苏省寄生虫病防治研究所、卫生部寄生虫病预防与控制技术重点实验室、江苏省寄生虫与媒介控制技术重点实验室,无锡214064

出　　处：《中国血吸虫病防治杂志》2016年第2期146-150,共5页Chinese Journal of Schistosomiasis Control

基　　金：江苏省科技厅能力提升项目(BM2015024-1);江苏省自然科学基金(BK20150001,BK2012106);国家自然科学基金(81271870,81401694);江苏省“六大人才高峰”项目(SWYY-004);无锡市卫生局科研项目(MS201528)

摘　　要：目的评价万孚疟原虫检测试剂盒（Pf-LDH/Pan-p LDH）对卵形疟原虫的检测效果,并分析原虫密度、疟原虫乳酸脱氢酶（p LDH）浓度和基因多态性等因素对检测结果的影响。方法按照万孚疟原虫检测试剂盒的操作说明书,对经PCR确认的100份卵形疟患者血样进行检测。采用显微镜镜检法计数患者血片的原虫密度,以ELISA法检测血样中的p LDH浓度,以PCR扩增卵形疟原虫LDH（Po-LDH）基因并测序,并分析上述3个因素对检出率的影响。结果万孚疟原虫检测试剂盒对上述100份卵形疟患者血样的总体检出率为70.0%（70/100）。当原虫密度≤500个/μl时,检出率为27.3%;原虫密度〉500个/μl时,检出率为75.0%～75.4%。当p LDH浓度较低时（A值≤0.100）,检出率为6.7%;p LDH达到一定浓度时（A值〉0.100）,检出率为95.1%～100%。Po-LDH基因序列分析结果显示所有卵形疟样本可分为2种亚型,分别为卵形疟原虫curtisi亚种（P.o.curtisi）和卵形疟原虫wallikeri亚种（P.o.wallikeri）。2种亚型的LDH基因同源性为97%,共有24个单核苷酸多态性（SNP）,其中3个SNPs为非同义突变,其余均为同义突变。2种亚型的LDH编码氨基酸序列同源性为99%,共有3个氨基酸的差异。万孚疟原虫检测试剂盒对P.o.curtisi的检出率为73.1%（38/52）,对P.o.wallikeri的检出率为66.7%（32/48）,两者差异无统计学意义（P〉0.05）。结论万孚疟原虫检测试剂盒对卵形疟原虫的检测效果优于大部分同类产品,其检出率与原虫密度、p LDH浓度关系密切,与p LDH基因多态性无关。Objective To evaluate the Wondfo Rapid Diagnostic Kit（Pf-LDH/Pan-p LDH）for detecting Plasmodium ovale and analyze the influence of parasitaemia,concentration and polymorphism of p LDH on the performances.Methods A total of100 blood samples from P.ovale patients confirmed by PCR were detected with the Wondfo Rapid Diagnostic Kit according to the manufacturers＇ instructions.The parasitaemia was determined by the microscopic examination.The concentration of p LDH was measured by ELISA tests.The LDH gene of P.ovale was amplified by PCR and sequenced.The influence of these three factors on the positive rate was analyzed.Results The overall positive rate of Wondfo Rapid Diagnostic Kit was 70.0%（70/100）.The positive rate was 27.3% for the samples with parasitaemia ≤ 500 parasites/μl and reached 75.0%-75.4% when parasitaemia 500 parasites/μl.The positive rate was 6.7% for samples with a low p LDH concentration（A values ≤ 0.100）and reached95.1%-100% at a high p LDH concentration（A values 0.100）.The results of sequence analysis indicated that all the samples could be divided into 2 types,P.o.curtisi and P.o.wallikeri.The gene homology of LDH between 2 types was 97%.There were24 single nucleotide polymorphism（s）（SNPs）between 2 types,while only 3 SNPs were non-synonymous mutations.The homology of LDH amino acid sequences between 2 types was 99%;only 3 amino acids were different.The positive rates for P.o.curtisi and P.o.wallikeri were 73.1%（38/52）and 66.7%（32/48）respectively;there was no statistically significant difference（P〉0.05）.Conclusion The Wondfo Rapid Diagnostic Kit（Pf-LDH/Pan-p LDH）performs better than most of the similar products for the detection of P.ovale,and the positive rates are closely related to the parasitaemia and concentration of p LDH,while no related to the polymorphism of p LDH gene.

关 键 词：卵形疟原虫 万孚疟原虫检测试剂盒 原虫密度 疟原虫乳酸脱氢酶 效果评价 影响因素 

分 类 号：R382.31[医药卫生—医学寄生虫学]