PCR扩增技术用于犬内脏利什曼病病原体kDNA的检测

PCR AMPLIFICATION OF LEISHMANIA DONOVANI kDNA FOR DIAGNOSIS OF CANINE VISCERAL LEISHMANIASIS

本文报导了用作者设计的一对寡核苷酸引物Ⅰ和Ⅱ以及文献报导的一对引物A、B检测内脏利什曼病病犬的骨髓、脾和外周血内的病原体kDNA,并和骨髓涂片检查利什曼原虫的结果进行了比较。共检测人工感染犬12只,骨髓涂片阳性者8例,其中PCR检测骨髓组织样品阳性者7例,检测脾组织样品阳性者3例;对照组犬脾、骨髓和外周血所有组织样品均为阴性。根据实验结果认为此法用于检测犬内脏利什曼病病原体kDNA有较好的效果。

Based on a set of oligonucleotide primers I and II and another set of primers A and B, poly- merase chain reaction was conducted to detect L. donovani kDNA in the spleen, bone marrow and peripheral blood cells from 12 dogs experimentally infected by inoculating L. donovani pro- mastigotes(MHOM/CH/90/SCW12)with different dosages and from 2 normal control dogs. A- mong the 12 infected dogs, 8 showed the presence of L. donovani amastigotes by bone marrow smear examination, of which the amplified products of PCR from bone marrow and spleen speci- mens were shown in 7 and 3 dogs respectively. No amplified product showed from spleen, bone marrow and peripheral blood cells from the normal dogs with the same way. It is suggested that PCR is a promising technique for diagnosis of visceral leishmaniasis.