【摘要】 采集大连大窑湾养殖区的长牡蛎(Crassostrea gigas),抽取其血淋巴液在显微镜下观察,观察到尼氏单孢子虫(Hap-losporidium nelsoni)原生质体的类似物。对抽取的血淋巴液进行总基因组DNA的提取,将扩增单孢子虫(Haplosporidiosis)SSU rDNA区域的引物对HAP-F2和HAP-R2进行改进,经过特异性试验证明其特异性很好,得到引物对HAP-F和HAP-R,应用此对引物进行聚合酶链式反应(PCR),结果扩增出239bp条带。将PCR产物进行测序,经序列比对分析确定为尼氏单孢子虫。同时,采用尼氏单孢子虫的特异性DNA探针MSX1347对抽取的长牡蛎血淋巴液进行原位杂交,结果也检测到了尼氏单孢子虫。更多还原

【Abstract】 Hemolymph extracted from Crassostrea gigas in Dayaowan Bay was examined by the microscope and Haplosporidium-like plasmodia stages were found in Hemolymph.DNA was extracted from the infected oyster Hemolymph for polymerase chain reaction(PCR) amplification of the haplosporidian.The primer HAP-F and HAP-R,which was modification of Hap-F2 and HAP-F2 previously reported,was specific to H.nelsoni by experiments and was applied to amplify short regions of small subunit ribosomal DNA(SSU rDNA) from H.nelsoni.PCR amplification of genomic DNA with primer HAP-R and HAP-F yielded the expected-sized product from the primer pair targeting the region for 239 bp.This PCR product was sequenced and it was identical to the corresponding SSU rDNA region of H.nelsoni.The Haplosporidium nelsoni-specific DNA probes MSX1347 hybridized with the C.gigas parasite.更多还原