摘要
目的 建立一种特异、敏感、简捷的PCR检测食蟹猴疟原虫（P.C）的方法。方法 检测样品来自人工接种感染P.C的阳性猴血膜及滤纸血斑。根据P.C SSUrRNA基因序列,设计P.C特异引物Pc1和Pc3,用PCR常规操作对P.C DNA模板扩增。同时用多对间日疟原虫（P.V）特异引物对照比较。结果 从 P.C血样中扩增出 425bp特定扩增带,而P.V红内期和子孢子、恶性疟原虫、伯氏疟原虫、人基因组DNA和健康猴血均未见扩增带;敏感度可检测 1.68 ×10-6的原虫感染水平。同时发现P.C对多对 P.V特异引物均出现阳性扩增带,以往一直未被注意。结论 该方法检测 P.C特异、敏感和简捷,提供了鉴别 P.V与 P.C的准确诊断。建议在 PCR的P.V引物设计和特异性的检测过程,把应用P.C DNA检测列为常规对照,以提高检测质量。
ve To establish a specific, sensitive and rapid polymerase chain reaction (PCR) method for detection of plasmodium cynomolgi (p.c) . Methods Blood film and filter spots were taken from monkeys artificially infected with p. c as samples. Primers Pc1 and Pc3 specific to p. c were designed based on ssUrRNA genetic sequence of p. c and the DNA template of p. c was amplified by conventional PCR. Meanwhile, Comparisons were made by using several pairs of Plasmoium vivax (p. v) primers. Results The expected amplified bands of 425bp were obtained from p.c samples, while no bands were identified from samples of p. v at blood stage and sporoites, plamsmodium falciarum, plasmodium bergeri, human genomic DNA and healthy monkey. The sensitivity reached a detecting level of 1.68 × 10-6 parasites. It was also observed that there were positive amplification bands in several specific p. v primers when detected by p. c samples, which has not been noticed. Conclusion This method is specific, sensitive and rapid in detecting p. c and it can differentiate the infection of p. v and p.c. It is suggested that the detection of p. c DNA be taken as conventional control in the process of designing p. v primers for specific detection by PCR so as to enhance the detection quanlity.
出处
《中国热带医学》
CAS
2002年第1期1-5,共5页
China Tropical Medicine
