两种检测方法在间日疟和卵形疟诊断中的应用分析
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摘要
目的分析疟原虫镜检和巢式PCR检测间日疟和卵形疟效果。方法收集2012～2016年输入性间日疟和卵形疟病例血样,巢式PCR检测疟原虫ssRNA基因,并与显微镜检结果进行比对。结果显微镜检和巢式PCR检测71份血样,间日疟、卵形疟、混合感染分别占74.6%、25.4%、0%和63.4%、29.6%、7%,符合率为81.7%;亚洲23份血样,镜检与巢式PCR均为间日疟,符合率为100%,巢式PCR检测非洲血样48份,间日疟、卵形疟和混合感染分别占45.8%、43.8%和10.4%,镜检与巢式PCR符合率为72.9%;巢式PCR检测26份卵形疟,经典curtisi、变种wallikeri和混合感染分别占80.8%、11.5%和7.7%,检出变种wallikeri总数占卵形疟19.2%。结论巢式PCR检测疟原虫虫种鉴别优于传统镜检法,可提高疟疾病例诊断水平。
Objective To assess the outcomes of applying microscopic examination and nested polymerase chain reaction( PCR) to detection of Plasmodium vivax( P. vivax) and Plasmodium ovale( P. ovale). Methods-Blood samples were collected from patients infected with P. vivax and P. ovale between 2012 and 2016. Nested PCR was performed to determine the Plasmodium ssRNA gene,and the results were compared with microscopic findings. Results A total of 71 blood samples were tested by microscopy and nested PCR. The two techniques revealed simple P. vivax,P. ovale and mixed infection by 74. 6%,25. 4% and 0%,and 63. 4%,29. 6% and7%,respectively. The coincidence rate was 81. 7%. In the 23 blood samples from Asia imported P. vivax cases,the coincidence rate was 100% by microscopic examination and nested PCR. Nested PCR revealed that P. vivax,P. ovale and mixed infection was 45. 8%,43. 8% and 10. 4%,respectively,in the 48 blood samples obtained from Africa imported cases. The coincidence rate was 72. 9% by two techniques. Twenty-six blood samples of P.ovale were tested by nested PCR,the results demonstrated that P. ovale curtisi( former classic type) and P. ovale wallikeri( former variant type) and mixed infection was 80. 8%,11. 5% and 7. 7%,respectively,and the total number of mutant wallikeri accounted for 19. 2% of P. ovale. Conclusion Nested PCR is superior to conventional microscopic examination in detection of plasmodium,and can improve the diagnosis of malaria.
Objective To assess the outcomes of applying microscopic examination and nested polymerase chain reaction( PCR) to detection of Plasmodium vivax( P. vivax) and Plasmodium ovale( P. ovale). Methods-Blood samples were collected from patients infected with P. vivax and P. ovale between 2012 and 2016. Nested PCR was performed to determine the Plasmodium ssRNA gene,and the results were compared with microscopic findings. Results A total of 71 blood samples were tested by microscopy and nested PCR. The two techniques revealed simple P. vivax,P. ovale and mixed infection by 74. 6%,25. 4% and 0%,and 63. 4%,29. 6% and7%,respectively. The coincidence rate was 81. 7%. In the 23 blood samples from Asia imported P. vivax cases,the coincidence rate was 100% by microscopic examination and nested PCR. Nested PCR revealed that P. vivax,P. ovale and mixed infection was 45. 8%,43. 8% and 10. 4%,respectively,in the 48 blood samples obtained from Africa imported cases. The coincidence rate was 72. 9% by two techniques. Twenty-six blood samples of P.ovale were tested by nested PCR,the results demonstrated that P. ovale curtisi( former classic type) and P. ovale wallikeri( former variant type) and mixed infection was 80. 8%,11. 5% and 7. 7%,respectively,and the total number of mutant wallikeri accounted for 19. 2% of P. ovale. Conclusion Nested PCR is superior to conventional microscopic examination in detection of plasmodium,and can improve the diagnosis of malaria.
引文
1雷蕾,买买提江·吾买尔,李志宏,等.输入性疟疾引起继发传播的风险评估研究进展.中国寄生虫学与寄生虫病杂志,2016,34(5):468-471.
2李黎,许国君,郁涛,等.2013年四川省疟疾流行病学特征分析.现代预防医学,2015,42(17):3097-3099.
3夏菁,蔡顺祥,林文,等.2010-2014年湖北省疟疾疫情流行病学分析.中国血吸虫病防治杂志,2016,28(3):247-251.
4中华人民共和国国家卫生和计划生育委员会.WS259-2015疟疾的诊断.北京:人民卫生出版社,2015.
5周水茂,杨燕,吴凯,等.非洲输入性恶性疟病例裂殖子表面蛋白1等位基因型检测.中国寄生虫学与寄生虫病杂志,2015,33(4):247-250.
6 Stephanie PJ,Norman JP,Maniphet VX,et al.PCR as a confirmatory technique for laboratory diagnosis of malaria.JClin Microbiol,2006,44(3):1087-1089.
7林耀莹,张山鹰,谢汉国,等.罕见输入性卵形疟原虫wallikeri亚种的PCR鉴定和测序分析.中国病原生物学杂志,2014,9(10):905-907.
8王亚丽,王煊,张彦平,等.2005~2013年全国本地感染与境外输入疟疾流行病学特征分析.中国媒介生物学及控制杂志,2015,26(2):120-125.
9江莉,王真瑜,张耀光,等.3种疟疾检测方法的应用分析.中国寄生虫学与寄生虫病杂志,2017,35(1):53-58.
10黄炳成,徐超,李瑾,等.巢式PCR技术在输入性卵形疟诊断中的应用研究.中国寄生虫学与寄生虫病杂志,2015,33(1):49-51.
11曹彩群,王伟明,丁桂生.2015年南通市疟疾血片复检质量分析.热带病与寄生虫学,2017,15(3):152-154.
12杨成运,钱丹,陈伟奇,等.河南省疟疾流行二类县专业技术人员疟原虫检测能力的评估分析.现代预防医学,2015,42(17):3165-3168.
13谭妍,李志峰,凌华,等.2015年重庆市3例输入性卵形疟原虫wallikeri亚种感染病例的实验室诊断.热带病与寄生虫学,2016,14(3):141-144.
14 Alemu A,Fuehrer HP,Getnet G,Plasmodium ovale curtisi and Plasmodium ovale wallikeri in North-West Ethiopia.Malar J,2013,12:346.
15辛辛,陆娟,郝莉鹏,等.上海浦东新区2014年亚信峰会公共卫生保障专题风险评估.公共卫生与预防医学,2014,25(4):4-7.
16徐俊芳,夏志贵,周晓农,等.我国2004-2013年疟疾发病率的时间趋势分析.中国寄生虫学与寄生虫病杂志,2017,35(2):114-118.
17雷露,刘学升,毛玲玲,等.3种实验室检测方法在疟疾诊断中的应用比较.中国媒介生物学及控制杂志,2015,26(2):206-207.
2李黎,许国君,郁涛,等.2013年四川省疟疾流行病学特征分析.现代预防医学,2015,42(17):3097-3099.
3夏菁,蔡顺祥,林文,等.2010-2014年湖北省疟疾疫情流行病学分析.中国血吸虫病防治杂志,2016,28(3):247-251.
4中华人民共和国国家卫生和计划生育委员会.WS259-2015疟疾的诊断.北京:人民卫生出版社,2015.
5周水茂,杨燕,吴凯,等.非洲输入性恶性疟病例裂殖子表面蛋白1等位基因型检测.中国寄生虫学与寄生虫病杂志,2015,33(4):247-250.
6 Stephanie PJ,Norman JP,Maniphet VX,et al.PCR as a confirmatory technique for laboratory diagnosis of malaria.JClin Microbiol,2006,44(3):1087-1089.
7林耀莹,张山鹰,谢汉国,等.罕见输入性卵形疟原虫wallikeri亚种的PCR鉴定和测序分析.中国病原生物学杂志,2014,9(10):905-907.
8王亚丽,王煊,张彦平,等.2005~2013年全国本地感染与境外输入疟疾流行病学特征分析.中国媒介生物学及控制杂志,2015,26(2):120-125.
9江莉,王真瑜,张耀光,等.3种疟疾检测方法的应用分析.中国寄生虫学与寄生虫病杂志,2017,35(1):53-58.
10黄炳成,徐超,李瑾,等.巢式PCR技术在输入性卵形疟诊断中的应用研究.中国寄生虫学与寄生虫病杂志,2015,33(1):49-51.
11曹彩群,王伟明,丁桂生.2015年南通市疟疾血片复检质量分析.热带病与寄生虫学,2017,15(3):152-154.
12杨成运,钱丹,陈伟奇,等.河南省疟疾流行二类县专业技术人员疟原虫检测能力的评估分析.现代预防医学,2015,42(17):3165-3168.
13谭妍,李志峰,凌华,等.2015年重庆市3例输入性卵形疟原虫wallikeri亚种感染病例的实验室诊断.热带病与寄生虫学,2016,14(3):141-144.
14 Alemu A,Fuehrer HP,Getnet G,Plasmodium ovale curtisi and Plasmodium ovale wallikeri in North-West Ethiopia.Malar J,2013,12:346.
15辛辛,陆娟,郝莉鹏,等.上海浦东新区2014年亚信峰会公共卫生保障专题风险评估.公共卫生与预防医学,2014,25(4):4-7.
16徐俊芳,夏志贵,周晓农,等.我国2004-2013年疟疾发病率的时间趋势分析.中国寄生虫学与寄生虫病杂志,2017,35(2):114-118.
17雷露,刘学升,毛玲玲,等.3种实验室检测方法在疟疾诊断中的应用比较.中国媒介生物学及控制杂志,2015,26(2):206-207.