长颈鹿血矛线虫ITS的PCR扩增与序列分析

PCR AMPIFICATION AND SEQUENCE ANALYSIS OF ITS RDNA OF HAEMONCHUS FROM A GIRAFFE IN CHINA

目的利用分子生物学方法对来自长颈鹿皱胃的血矛线虫进行虫种鉴定。方法对样品XM9和XM11的核糖体DNA内转录间隔区(ITS-1、5.8 S、ITS-2)进行PCR扩增及序列分析,并与GenBank公布的血矛线虫(Hae-monchus)相应序列进行比较。结果来自长颈鹿皱胃的2条血矛线虫具有相同的ITS序列,5.8 S与ITS-1分别为153 bp、404 bp,与GenBank分布的捻转血矛线虫序列是一致的。ITS-2序列为231 bp,第753位是一个多态位点,该序列与来自国外的H.contortus,H.placei,H.longistipes存在0-18个碱基差异。结论来自长颈鹿的血矛线虫是捻转血矛线虫。

Objective To determine the specific status of two Haemonchus specimens （XM9 and XM11 ） collected from a giraffe in Guangdong Province of China. Methods The internal transcribed spacers（ITS-1,5.8 S, ITS-2） of ribosomal DNA were amplified by PCR and the amplicons were sequenced. The sequences were compared with those of H. contortus, H. placei, and H. longistipes, available in GenBank. Results The results revealed that XM9 and XM11 had the same ITS sequences, their lengths being 403 bp（ITS-1 ）, 153 bp（5.8 S） and 231 bp（ITS-2） respectively. The sequences of 5.8 S and ITS-1 were consistent with those of H. contortus from Australia, and the 753th base in ITS was a polymorphic site. The ITS-2 sequence differed to that of H. contortus, H. placei, and H. longistipes by 0 to 18 bases. Conclusion These results indicated that XM9 and XMll represented H. contortus.