Wu Z, Nagano I, Takahashi Y
Department of Parasitology, Gifu University School of Medicine, Japan.
Parasitology. 1998 Aug;117 ( Pt 2):173-83. doi: 10.1017/s0031182098002881.
Diagnostic PCR primers for Trichinella were constructed. Twelve pairs of primers were designed based on the sequences of random amplified polymorphic DNA, and 4 pairs of primers were designed based on the reported sequences of complementary DNA encoding excretory-secretory glycoproteins. With these primers, 31 samples of DNA from different strains of Trichinella including 5 species (Trichinella spiralis, Trichinella britovi, Trichinella nativa, Trichinella nelsoni and Trichinella pseudospiralis) and 3 phenotypes of uncertain taxonomic level (Trichinella T5, T6 and T8) were tested with PCR. Genus Trichinella can be identified by 4 different primer pairs (SB147D, SB372A, SB153, or Ts43). Trichinella spiralis can be identified by the presence of a 673 bp amplicon in PCR with the primer pair SB147B. Trichinella nelsoni can be identified using primer pair SB147F or by the presence of 673 bp and ca. 380 bp amplicon in PCR with the primer pair SB147B. Trichinella pseudospiralis can be identified by 2 primer pairs (SB147E or SB372B). Trichinella T5 can be identified by the primer pair SB147G. Trichinella T8 can be identified by its positivity by the primer pair SB147C and its negativity by the primer pair SB372C. A group of Trichinella species (T. britovi, T. nativa and Trichinella T6) can be identified by the primer pair SB372C.
构建了旋毛虫诊断性PCR引物。基于随机扩增多态性DNA序列设计了12对引物,基于已报道的编码排泄分泌糖蛋白的互补DNA序列设计了4对引物。利用这些引物,对31份来自不同旋毛虫菌株的DNA样本进行了PCR检测,这些菌株包括5个种(旋毛形线虫、布氏旋毛虫、本地毛形线虫、纳氏旋毛虫和伪旋毛虫)以及3种分类地位不确定的表型(旋毛虫T5、T6和T8)。旋毛虫属可通过4种不同的引物对(SB147D、SB372A、SB153或Ts43)进行鉴定。在使用引物对SB147B进行的PCR中,若出现673 bp扩增子,则可鉴定为旋毛形线虫。使用引物对SB147F或在使用引物对SB147B进行的PCR中出现673 bp和约380 bp扩增子,可鉴定为纳氏旋毛虫。伪旋毛虫可通过2对引物(SB147E或SB372B)进行鉴定。旋毛虫T5可通过引物对SB147G进行鉴定。旋毛虫T8在使用引物对SB147C时呈阳性,在使用引物对SB372C时呈阴性,据此可对其进行鉴定。一组旋毛虫种类(布氏旋毛虫、本地毛形线虫和旋毛虫T6)可通过引物对SB372C进行鉴定。