{"title":"牛血清大肠杆菌感染特异性DNA探针的研究。","authors":"N Kajiwara, R Kirisawa, M Onuma, Y Kawakami","doi":"10.1292/jvms1939.52.1199","DOIUrl":null,"url":null,"abstract":"<p><p>A simple procedure was developed for detection of Theileria sergenti infection on the basis of hybridization of parasite DNA with a specific probe. A genomic DNA library of T. sergenti constructed in pUC-18 was screened to detect clones containing the parasite's DNA sequences by colony and Southern hybridizations. Two positive DNA inserts were purified from the recombinant plasmids and used as probes labelled with 32P or non-isotopic reagent, biotin-11-dUTP. 32P-radiolabelled and non-radioactive probes appear to be sensitive enough to detect 15 pg (equivalent to 1,200 parasites) and 125 pg (equivalent to 10,000 parasites) of purified T. sergenti DNA, and in diluted T. sergenti-infected red blood cells, they are able to detect 8,000 parasites and 16,000 parasites, respectively.</p>","PeriodicalId":19620,"journal":{"name":"Nihon juigaku zasshi. The Japanese journal of veterinary science","volume":"52 6","pages":"1199-204"},"PeriodicalIF":0.0000,"publicationDate":"1990-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1292/jvms1939.52.1199","citationCount":"11","resultStr":"{\"title\":\"Specific DNA probe for the detection of Theileria sergenti infection in cattle.\",\"authors\":\"N Kajiwara, R Kirisawa, M Onuma, Y Kawakami\",\"doi\":\"10.1292/jvms1939.52.1199\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>A simple procedure was developed for detection of Theileria sergenti infection on the basis of hybridization of parasite DNA with a specific probe. A genomic DNA library of T. sergenti constructed in pUC-18 was screened to detect clones containing the parasite's DNA sequences by colony and Southern hybridizations. Two positive DNA inserts were purified from the recombinant plasmids and used as probes labelled with 32P or non-isotopic reagent, biotin-11-dUTP. 32P-radiolabelled and non-radioactive probes appear to be sensitive enough to detect 15 pg (equivalent to 1,200 parasites) and 125 pg (equivalent to 10,000 parasites) of purified T. sergenti DNA, and in diluted T. sergenti-infected red blood cells, they are able to detect 8,000 parasites and 16,000 parasites, respectively.</p>\",\"PeriodicalId\":19620,\"journal\":{\"name\":\"Nihon juigaku zasshi. The Japanese journal of veterinary science\",\"volume\":\"52 6\",\"pages\":\"1199-204\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1990-12-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1292/jvms1939.52.1199\",\"citationCount\":\"11\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Nihon juigaku zasshi. The Japanese journal of veterinary science\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1292/jvms1939.52.1199\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Nihon juigaku zasshi. The Japanese journal of veterinary science","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1292/jvms1939.52.1199","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 11
摘要
建立了一种简单的方法,在寄生虫DNA与特异性探针杂交的基础上检测血吸虫感染。利用pcc -18构建的sergenti基因组DNA文库,通过集落杂交和Southern杂交检测含有该寄生虫DNA序列的克隆。从重组质粒中纯化两个阳性DNA插入片段,用32P或非同位素试剂生物素-11- dutp标记探针。32p放射性标记探针和非放射性探针似乎足够敏感,可以检测到15pg(相当于1200个寄生虫)和125pg(相当于10000个寄生虫)纯化的sergenti T. DNA,在稀释的sergenti T.感染的红细胞中,它们分别能够检测到8000个寄生虫和16000个寄生虫。
Specific DNA probe for the detection of Theileria sergenti infection in cattle.
A simple procedure was developed for detection of Theileria sergenti infection on the basis of hybridization of parasite DNA with a specific probe. A genomic DNA library of T. sergenti constructed in pUC-18 was screened to detect clones containing the parasite's DNA sequences by colony and Southern hybridizations. Two positive DNA inserts were purified from the recombinant plasmids and used as probes labelled with 32P or non-isotopic reagent, biotin-11-dUTP. 32P-radiolabelled and non-radioactive probes appear to be sensitive enough to detect 15 pg (equivalent to 1,200 parasites) and 125 pg (equivalent to 10,000 parasites) of purified T. sergenti DNA, and in diluted T. sergenti-infected red blood cells, they are able to detect 8,000 parasites and 16,000 parasites, respectively.