Scott J C, McManus D P
Molecular Parasitology Unit, Queensland Institute of Medical Research, Brisbane, Australia.
Trop Med Parasitol. 1994 Mar;45(1):1-4.
The random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), also called arbitrary priming PCR (AP-PCR), is a DNA polymorphism assay based on the amplification of genomic DNA using a single oligonucleotide primer of arbitrary nucleotide sequence. Using carefully controlled conditions, the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) proved useful for distinguishing Echinococcus species and genetically distinct strains of Echinococcus granulosus. Amongst twenty random 10 mer oligonucleotide primers tested, 4 (5'-GGGAATTCGG-3', 5'-GGCTGCAGAA-3', 5'-GGAGTACTGG-3', 5'-CCTCTAGACC-3') were most suitable for producing complex but characteristic patterns for discrimination purposes. Although the method amplifies vertebrate DNA, the resulting patterns do not interfere with the identification of Echinococcus isolates. Consequently, the technique can still be used effectively to distinguish Echinococcus species and strains even if host contamination occurs during the course of parasite collection. The approach complements several other methods, based on direct DNA examination, recently developed in our laboratory, which also allow unambiguous discrimination of Echinococcus genotypes.
随机扩增多态性DNA聚合酶链反应(RAPD-PCR),也称为任意引物PCR(AP-PCR),是一种基于使用具有任意核苷酸序列的单个寡核苷酸引物扩增基因组DNA的DNA多态性检测方法。在精心控制的条件下,随机扩增多态性DNA聚合酶链反应(RAPD-PCR)被证明可用于区分棘球绦虫属物种以及细粒棘球绦虫的遗传不同菌株。在所测试的20个随机10聚体寡核苷酸引物中,4个(5'-GGGAATTCGG-3'、5'-GGCTGCAGAA-3'、5'-GGAGTACTGG-3'、5'-CCTCTAGACC-3')最适合用于产生复杂但具有特征性的图谱以进行鉴别。尽管该方法会扩增脊椎动物的DNA,但所产生的图谱不会干扰棘球绦虫分离株的鉴定。因此,即使在寄生虫采集过程中发生宿主污染,该技术仍可有效地用于区分棘球绦虫属的物种和菌株。该方法补充了我们实验室最近开发的其他几种基于直接DNA检测的方法,这些方法也能够明确区分棘球绦虫的基因型。