Bendezu Ivan F, Slater John W, Carney Brian F
Science Department, Letterkenny Institute of Technology, Port Road, Letterkenny, County Donegal, Ireland.
Mar Biotechnol (NY). 2005 Nov-Dec;7(6):687-96. doi: 10.1007/s10126-004-0124-y. Epub 2005 Oct 3.
Two molecular protocols for the identification of mussel and scallop have been developed using specific primers targeting the mitochondrial 16S ribosomal DNA gene and the nuclear 18S ribosomal DNA gene. Primers for the mitochondrial 16S ribosomal DNA gene in multiplex polymerase chain reaction (PCR) protocols yielded diagnostic DNA fragments for the mussels Mytilus edulis, Mytilus galloprovincialis, and the hybrid Mytilus edulis/galloprovincialis (335 bp), the king scallop Pecten maximus (382 bp) and the black scallop Mimachlamys varia (398 bp). DNA from the queen scallop Aequipecten opercularis showed no consistent PCR amplification of the 16S rDNA gene. Primers for the nuclear 18S rDNA gene in standard PCR protocols yielded similar-sized, diagnostic DNA fragments (approx. 190 bp) for the mussels Mytilus edulis, Mytilus galloprovincialis, and the hybrid Mytilus edulis/galloprovincialis, the king scallop Pecten maximus, the black scallop Mimachlamys varia, and the queen scallop Aequipecten opercularis. Both protocols have been tested with Mytilus spp., P. maximus, and 6 other bivalve species from a wide range of locations in Irish and European waters. Cross reaction of the specific primers with DNA template from any of the 6 other bivalve species was not observed. Rapid DNA extraction using FTA Card technology and the16S rDNA primers allowed for the detection of at least 10 mussel larvae in a subsample of natural plankton.
已开发出两种用于鉴定贻贝和扇贝的分子方法，使用针对线粒体16S核糖体DNA基因和核18S核糖体DNA基因的特异性引物。多重聚合酶链反应（PCR）方法中用于线粒体16S核糖体DNA基因的引物，产生了可用于诊断的DNA片段，这些片段来自贻贝紫贻贝、地中海贻贝以及紫贻贝/地中海贻贝杂交种（335 bp）、王扇贝（382 bp）和黑扇贝（398 bp）。皇后扇贝的DNA在16S rDNA基因的PCR扩增中未表现出一致的扩增结果。标准PCR方法中用于核18S rDNA基因的引物，为紫贻贝、地中海贻贝以及紫贻贝/地中海贻贝杂交种、王扇贝、黑扇贝和皇后扇贝产生了大小相似的诊断性DNA片段（约190 bp）。这两种方法均已在贻贝属、王扇贝以及来自爱尔兰和欧洲水域广泛地点的其他6种双壳类物种上进行了测试。未观察到特异性引物与其他6种双壳类物种中任何一种的DNA模板发生交叉反应。使用FTA卡技术和16S rDNA引物进行快速DNA提取，能够在天然浮游生物的子样本中检测到至少10只贻贝幼虫。
