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目的建立粪便中隐孢子虫的提取与检测技术,分析腹泻病人粪便中隐孢子虫的感染情况,为该病的诊断、防治提供科学依据。方法选择经病原学检测已经确诊的5份隐孢子虫阳性和4份阴性粪便,分别采用FTA卡和商业化DNA抽提试剂盒抽提粪便样本中DNA,经荧光PCR法进行扩增,比较核酸提取效果;同时用直接涂片法、改良抗酸染色法及荧光PCR法对63例腹泻病人隐孢子虫感染情况进行粪样检测分析。结果应用FTA卡抽提DNA效果优于应用商业化DNA抽提试剂盒抽提DNA。同时对63份腹泻病人粪便运用直接涂片法、改良抗酸染色法及荧光PCR法都未检出隐孢子虫。结论应用FTA卡进行粪便样本中隐孢子虫DNA的抽提方法具有简单有效、快速灵敏、准确可靠等特点,可以应用于粪便中隐孢子虫的检测,同时可联合多种检测手段来提高腹泻病人原虫感染的检出率。
Objective To establish the extraction and detection of Cryptosporidium in feces and to analyze the infection of Cryptosporidium in feces of diarrhea patients so as to provide a scientific basis for the diagnosis and prevention of this disease. Methods Five pathogenically confirmed positive samples of Cryptosporidium positive and four negative samples of stools were selected. DNA was extracted from the stool samples by FTA card and commercial DNA extraction kit respectively. The amplified DNA was amplified by fluorescence PCR. The results of stool samples were also analyzed by direct smear method, modified acid-fast staining method and fluorescent PCR method in 63 cases of diarrhea patients with cryptosporidiosis. Results The DNA extraction using FTA card was better than DNA extraction kit using commercial DNA extraction kit. At the same time, Cryptosporidium was not detected in 63 diarrhea patients by direct smear method, modified acid-fast staining method and fluorescent PCR method. Conclusions The extraction of Cryptosporidium DNA from stool samples by FTA card is simple, effective, rapid and sensitive, accurate and reliable. It can be applied to the detection of cryptosporidium in stool samples. At the same time, a combination of detection methods can be used to improve diarrhea patients Protozoan infection detection rate.