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应用一种新的非放射标记物地高辛配基(DIG)制备黄疸出血群赖型017株钩端螺旋体DNA(017DNA)探针,与~(32)P和光敏生物素标记017DNA探针对照打点杂交检测钩端螺旋体DNA。结果,上述三种探针均能检测到不同群型的问号状钩体,DIG探针的敏感性为0.1pg,~(32)P和光敏生物素探针的敏感性分别为1 pg和10pg。以上探针均不能检测双曲钩体patoc型Patocl株、细螺旋体illini型3065株,与其它致病菌及人白细胞DNA也无明显杂交信号。DIG标记017DNA探针原位杂交检测感染017株钩体的豚鼠系列组织切片和血浆涂片,以对比度极大的着色反应,清晰地显示各组织和血浆中的钩体形态。细螺旋体illini型3055探针和HBV-DNA探针不能检测到上述标本中的钩体.
A new non-radiolabeled digoxigenin (DIG) was used to prepare 017 leptospira DNA (017DNA) probes of jaundice hemorrhagic group, which were compared with ~ (32) P and photobiotin labeled 017DNA probe RBI hybridization detection of leptospiral DNA. As a result, each of the above-mentioned three types of probes could detect different types of tick-like bodies, the sensitivity of the DIG probe was 0.1 pg, the sensitivity of the (32) P and the photosensitizer probe was 1 pg and 10 pg . None of the above probes could detect patoc-type patocl strains of Leptospira and illini-type 3065 strains of Leptospira, which showed no obvious hybridization signal with other pathogenic bacteria and human leukocyte DNA. DIG-labeled 017 DNA probe in situ hybridization was used to detect the tissue sections and plasma smears of the guinea pigs infected with 017 strains of leptospirosis. The results showed that the morphology of the hooks in each tissue and plasma was clearly shown by the contrasting coloration reaction. The leptospira illini-type 3055 probe and the HBV-DNA probe failed to detect the leptospira in the above specimens.