Leng Z, Riley D E, Berger R E, Krieger J N, Roberts M C
Department of Pathobiology, University of Washington, Seattle 98195, USA.
J Antimicrob Chemother. 1997 Oct;40(4):551-9. doi: 10.1093/jac/40.4.551.
We tested 34 American Type Culture Collection (ATCC) and 168 clinical bacterial isolates, from the human urogenital and oral tracts and streptococci isolated from cows with mastitis, for the presence of the tetQ gene using a polymerase chain reaction (PCR) assay and DNA-DNA hybridization. The identities of PCR products were confirmed by Southern blot hybridization of whole-cell DNA. Eleven of the ATCC strains were positive for tetQ, including five Bacteroides spp., five Prevotella spp. and a single isolate of Mitsuokella multiacidus. Twenty-eight (29%) of the 95 clinical Gram-negative isolates carried the tetQ gene, while eight (11%) of the 73 clinical Gram-positive isolates carried the tetQ gene. This is the first description of tetQ in Gram-positive species. All isolates except one Peptostreptococcus sp. carried tetQ integrated into the chromosome. The tetQ gene could be transferred from Prevotella bivia, Bacteroides ovatus, Bacteroides fragilis, Bacteroides vulgatus and Bacteroides distasonis into an Enterococcus faecalis recipient at frequencies of 10(-7)-10(-9) per recipient. In contrast, tetQ failed to transfer from two isolates of Prevotella intermedia, two isolates of Porphyromonas gingivalis, one isolate of Mobiluncus curtisii and one isolate of Peptostreptococcus sp. The latter two are Gram-positive species. The PCR assay was used to screen 198 proteinase K-treated biopsies of prostate, periprostate and bladder from 84 men with prostatitis. Thirty-four (40%) of the patients had one or more positive samples, suggesting that the PCR assay could be of value in screening patient material directly for the presence of bacteria.
我们使用聚合酶链反应(PCR)检测法和DNA-DNA杂交技术,检测了34株美国典型培养物保藏中心(ATCC)菌株以及168株临床分离细菌,这些临床分离细菌来自人类泌尿生殖道和口腔,以及从患乳腺炎奶牛分离出的链球菌,以检测tetQ基因的存在情况。通过全细胞DNA的Southern印迹杂交确认了PCR产物的身份。11株ATCC菌株tetQ呈阳性,包括5株拟杆菌属、5株普雷沃菌属和1株多酸 Mitsuokella 菌的分离株。95株临床革兰氏阴性分离株中有28株(29%)携带tetQ基因,而73株临床革兰氏阳性分离株中有8株(11%)携带tetQ基因。这是首次在革兰氏阳性菌中描述tetQ基因。除1株消化链球菌属细菌外,所有分离株携带的tetQ基因均整合到染色体中。tetQ基因可以以前列腺双栖普雷沃菌、卵形拟杆菌、脆弱拟杆菌、普通拟杆菌和狄氏拟杆菌为供体,以10(-7)-10(-9)的频率转移到粪肠球菌受体中。相比之下,tetQ未能从2株中间普雷沃菌、2株牙龈卟啉单胞菌、1株柯氏动弯杆菌和1株消化链球菌属细菌中转移。后两者为革兰氏阳性菌。PCR检测法用于筛查84例前列腺炎男性患者的198份经蛋白酶K处理的前列腺、前列腺周围组织和膀胱活检样本。34例(40%)患者有一个或多个阳性样本,这表明PCR检测法在直接筛查患者材料中细菌的存在方面可能具有价值。