Avidor B, Kletter Y, Abulafia S, Golan Y, Ephros M, Giladi M
The Bernard Pridan Laboratory for Molecular Biology of Infectious Diseases, Ichilov Hospital, Tel-Aviv Sourasky Medical Center, Israel.
J Clin Microbiol. 1997 Aug;35(8):1924-30. doi: 10.1128/jcm.35.8.1924-1930.1997.
Amplification of Bartonella henselae DNA has been proposed as a diagnostic test for cat scratch disease (CSD). The sensitivities of the following three PCR assays were compared. PCR/rRNA with universal primers amplifies part of the 16S rRNA gene, followed by hybridization with a specific B. henselae probe; PCR/CS and PCR/HSP amplify portions of the gltA and the htrA genes, respectively, each followed by restriction fragment length polymorphism analysis. The threshold of detection of B. henselae DNA in pus was 10(-4), 10(-3), and 10(-2) ng for PCR/rRNA, PCR/CS, and PCR/HSP, respectively. By these three assays, B. henselae DNA was detected in 100, 94, and 69% of 32 pus and lymph node specimens from CSD patients, respectively. The similar sensitivities of the PCR/rRNA and the PCR/CS assays for detecting B. henselae DNA in clinical specimens are in contrast to the 10-fold difference in sensitivities in favor of PCR/rRNA demonstrated with purified B. henselae DNA in sterile pus, suggesting that in the majority of cases, the bacterial load in clinical specimens is large enough to be identified by the PCR/CS assay. A two-step approach is suggested to achieve maximal sensitivity for detecting B. henselae in clinical specimens: initial testing by PCR/CS (which does not require hybridization), followed by PCR/rRNA with PCR/CS-negative specimens when CSD is strongly suspected.
已有人提出将汉赛巴尔通体DNA扩增作为猫抓病（CSD）的诊断检测方法。比较了以下三种聚合酶链反应（PCR）检测方法的灵敏度。使用通用引物的PCR/rRNA可扩增16S rRNA基因的一部分，随后与特异性汉赛巴尔通体探针杂交；PCR/CS和PCR/HSP分别扩增gltA基因和htrA基因的部分片段，每种方法之后都进行限制性片段长度多态性分析。对于PCR/rRNA、PCR/CS和PCR/HSP，在脓液中检测到汉赛巴尔通体DNA的阈值分别为10^(-4)、10^(-3)和10^(-2)纳克。通过这三种检测方法，在来自CSD患者的32份脓液和淋巴结标本中，分别有100%、94%和69%检测到了汉赛巴尔通体DNA。PCR/rRNA和PCR/CS检测方法在临床标本中检测汉赛巴尔通体DNA的灵敏度相似，这与在无菌脓液中使用纯化的汉赛巴尔通体DNA所显示的灵敏度相差10倍且有利于PCR/rRNA的情况形成对比，这表明在大多数情况下，临床标本中的细菌载量足以通过PCR/CS检测方法进行鉴定。建议采用两步法以实现临床标本中检测汉赛巴尔通体的最大灵敏度：首先通过PCR/CS进行初始检测（不需要杂交），当强烈怀疑为CSD时，对PCR/CS阴性的标本再进行PCR/rRNA检测。
