用虫种特异性线粒体基因序列鉴定2例疑似曼氏裂头蚴感染无虫病理标本

Use specific sequences of Mitochondrial gene to indentify two cases suspected plerocercoid infection pathological specimens with no worms

目的探讨采用线粒体细胞色素C氧化酶亚基1(CO1)基因特异性序列鉴定临床无虫病理标本中裂头蚴感染的可靠性。方法应用免疫曼氏裂头蚴的兔血清检测病理切片中裂头蚴的抗原,有2例呈阳性,疑似裂头蚴感染的无虫病例,按照DNA提取试剂盒说明书提取核酸,用曼氏裂头蚴线粒体CO1基因特异性序列的两对引物作扩增。结果经PCR 2次扩增后得到目的序列,电泳条带出现在预期的位置。2例病理标本扩增的核苷酸序列与已知基因片断序列比对,引物F650/R800扩增条带符合率分别为100%和97%,引物F965/R1120扩增条带符合率均为100%。结论用裂头蚴线粒体CO1特异性基因引物作重复PCR扩增,对鉴定无虫病理标本曼氏裂头蚴感染有诊断意义。

Purpose To explore the reliability of the specific sequences of the mitochondrial cytochrome Coxidase 1 （ CO1 ） gene which is used in indentifying the infection of sparganum in clinical pathological specimens with no worms. Methods Serum of rabbits immu- nized with the sparganum was used to examine the antigen from the sparganum. In the pathological slices two cases of the pathological sections showed positive. Finally, the two no-worm cases suspected as sparganosises were chosen for molecular experiment. Following the handbook of QIAamp DNA FFPE Tissue kit to extract DNA, two pairs of specific primers were used to amplify two partial sequences in the mitochondrial cytochrome CO1 gene. Results The targeted sequences were obtained through repeated PCR. The electrophoretic bands met the expectation. Blasted with the known sequences of sparganum in the pubmed, the accuracy of the sequences which was amplified by the pairs primes of F650/R800 in two chosen cases were 100% and 97% , respectively. And the other pairs primes of F965/R1120 were 100% and 100% , respectively. Conclusion Repeating PCR by the specific primers of CO1 gene of spaganum has diagnostic value in indentifying the pathological specimens of no worms.