摘要
通过建立能对石蜡切片中鹅细小病毒(GPV)核酸进行定位的原位PCR方法,为GPV在鹅体内的定位、致病机理研究等提供有效的试验手段。根据GPV的VP3基因序列设计PCR引物和寡核苷酸探针,以GPV感染鹅肝脏和空肠组织石蜡标本制作切片,经蛋白酶K消化、原位PCR扩增和碱性磷酸酶标记的寡核苷酸探针原位杂交,建立了检测石蜡标本中GPV的间接原位PCR方法并应用于自然感染GPV高免血清紧急免疫后鹅肝脏和空肠组织临床病料检测。结果显示间接原位PCR对人工感染GPV死亡鹅肝脏和空肠的石蜡标本检测结果为阳性,而鹅病毒性肝炎、鹅多杀性巴氏杆菌病、鹅沙门菌病和鹅大肠杆菌病死亡鹅肝脏的石蜡标本检测结果为阴性;间接原位PCR对自然感染GPV高免血清紧急免疫后第6天的鹅肝脏和空肠组织检测20个样本中,空肠组织有8个呈阳性,肝脏组织有4个阳性结果均为阳性,阳性细胞有空肠上皮细胞、肝窦上皮细胞等。
A pair of primers and an oligonucleotide probe were designed by primer 5.0 software according to the VP3 sequence of goose parvovirus in GenBank. The indirect in situ polymerase chain reaction was developed for detecting goose parvovirus in paraffin sections, which were used to test goose parvovirus in liver and jejunum of goose after immediate vaccination. The positive results were gained by indirect ISPCR for detecting parvovirus in the liver and jejunum paraffin tissues of geese infected by GPV. In comparison, out of 20 samples, Pasteurella multocida,viral hepatitis virus, Salmonella typhimurium, E. coli were detected in 8 samples of jejunum and 4 samples of liver tissues from the geese infected with GPV. The parvovirus could be detected from jejunum epithelial cells and liver sinusoidal endothelial cells by indirect IS-PCR.
出处
《中国兽医杂志》
CAS
北大核心
2011年第12期10-12,I0001,共4页
Chinese Journal of Veterinary Medicine
基金
吉林省教育厅青年基金(2010322)
吉林农业大学青年启动基金(2009083)
