多重巢式聚合酶链反应在疟疾混合感染检测与分型中的应用

发布时间：2018-04-23 04:35

本文选题：疟原虫 + 巢式聚合酶链反应　；参考：《东南大学学报(医学版)》2015年02期

【摘要】：目的:应用能够同时检测恶性疟(P.falciparum)、间日疟(P.vivax)、卵形疟(P.ovale)、三日疟(P.malarie)及其混合感染的多重巢式PCR方法,对疑似疟疾混合感染样本进行检测,评价其在疟原虫混合感染及分型中的应用价值,旨在为疟疾流行病学调查、疟疾混合感染的确诊提供一种快速高效、特异性强、灵敏度高的实验室诊断方法,为传统的镜检法提供有效的补充。方法:根据恶性疟原虫、间日疟原虫、卵形疟原虫和三日疟原虫的18SSU rRNA基因序列设计5对引物(1对通用引物、4对特异性引物),以等比例混合的4种疟原虫核酸为模板,建立疟疾多重巢式PCR检测方法,并应用该法对28份疑似疟疾混合感染样本进行检测及测序验证,将其结果与镜检法比较。结果:28份疑似混合感染样本中混合感染样品数为6例,其中恶性疟和间日疟混合感染样品5例,恶性疟和卵形疟混合感染样品1例。结论:本研究建立的多重巢式PCR检测方法能够同时用于恶性疟、间日疟、卵形疟、三日疟及其混合感染样本的检测,在疟疾混合感染的确诊与分型上较镜检法有明显的优势和更高的应用价值。
[Abstract]:Objective: to use a multiplex PCR method to detect P. falciparum, P. vivaxi, P. ovalei, P. malarie and their co-infection, and to detect the suspected malaria mixed infection samples, which can be used to detect P. falciparum, P. vivaxi, P. ovalei, P. malarie and their co-infection simultaneously, and to detect the combined infection of P. falciparum and P. vivaxi. The purpose of this study was to provide a rapid, efficient, specific and sensitive laboratory diagnostic method for malaria epidemiological investigation and diagnosis of malaria mixed infection. It provides an effective supplement for the traditional mirror examination method. Methods: according to the 18SSU rRNA gene sequence of Plasmodium falciparum, Plasmodium vivax, Plasmodium ovum and Plasmodium falciparum, five pairs of primers, one pair of universal primers, four pairs of specific primers were designed. A multiplex nested PCR method for malaria detection was established. 28 samples of suspected malaria mixed infection were detected and sequenced. The results were compared with the microscopic method. Results there were 6 cases of mixed infection among 28 suspected mixed infection samples, including 5 cases of falciparum malaria and vivax malaria mixed infection sample and 1 case of falciparum malaria and ovum malaria mixed infection sample. Conclusion: the multiplex nested PCR assay developed in this study can be used for the detection of falciparum malaria, vivax malaria, ovum malaria, 3-day malaria and their mixed infection samples at the same time. In the diagnosis and typing of malaria mixed infection, it has obvious advantages and higher application value than the method of microscopic examination.
【作者单位】：河南出入境检验检疫局;
【基金】：国家质检总局科研基金资助项目(2011IK131)
【分类号】：R531.3

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