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目的了解中国TTV基因分型状况。方法采用套式PCR检测D8、 D10、 D22、 D25、 D29血清中TTV DNA,限制性内切酶和PCR产物直接测序技术进行酶切和序列分析。结果 N22~ D8存在 BstUI特异性切点, G1b~D10无 BstUI切点,有EcoRI切点,G2a、D25、D29序列均存在Hae和Tsp509切点,G2b-D22除存在HaeⅢ切点之外还具有MboⅠ切点。Pat3~TTh6仅有Hae Ⅲ切点。结论AB008394~D8具有相同的切点可能均属la型,G1b-Jcase5切点相同可能为1b型,TTh1~1和TTh1~2可能属1型中的其它亚型。D25~D29酶切点相同为2a型,G2b~D22切点相同为2b型,Pat3~TTh6切点相同可能为2型中的其它亚型。
Objective To understand the status of TTV genotyping in China. Methods TdDNA of D8, D10, D22, D25 and D29 were detected by nested PCR and restriction endonucleases and direct sequencing of PCR products were digested and sequenced. Results There were BstUI specific cleavage sites on N22 ~ D8, no BstUI cleavage sites on G1b ~ D10, EcoRI cleavage sites, Hae and Tsp509 cleavage sites on G2a, D25 and D29 sequences, G2b-D22 had HaeⅢ cleavage sites Mbo I cut point. Pat3 ~ TTh6 only Hae Ⅲ cut point. Conclusion The same point of cleavage between AB008394 and D8 may belong to la type. The same point of G1b-Jcase5 may be type 1b, and TTh1-1 and TTh1-2 may belong to other subtypes of type 1. D25 ~ D29 digestion point the same as 2a type, G2b ~ D22 same point of the same type 2b, Pat3 ~ TTh6 same point may be other subtypes of type 2.