Schläpfer G, Cherry J D, Heininger U, Uberall M, Schmitt-Grohé S, Laussucq S, Just M, Stehr K
Abt. med. Mikrobiologie, Universitätskinderklinik, Basel, Switzerland.
Pediatr Infect Dis J. 1995 Mar;14(3):209-14. doi: 10.1097/00006454-199503000-00008.
The polymerase chain reaction (PCR) was recently added to conventional culture and serology for the diagnoses of Bordetella pertussis infection in a large vaccine efficacy trial in Germany. In vaccinees or family members who had illnesses with cough, two nasopharyngeal swabs (calcium alginate for culture and Dacron for PCR) were taken and initial and follow-up clinical data were obtained. PCR was done using oligonucleotide primers PTp1 and PTp2 which amplify a 191-base pair DNA fragment of pertussis toxin operon. From December, 1993, to May, 1994, 555 pairs of swabs were processed; 28 grew B. pertussis and 9 grew B. parapertussis. Twenty-six of the 28 subjects with B. pertussis-positive cultures also had positive PCR results as did one of the 9 B. parapertussis cases and 82 additional samples were positive by PCR. PCR increased the identification of subjects with B. pertussis infections by almost 4-fold. Clinical characteristics were analyzed by laboratory category (Group 1, 28 culture-positive; Group 2, 82 culture-negative, PCR-positive; and Group 3, 436 culture- and PCR-negative). Group 1 subjects were more likely to have a diagnosis of definite or probable pertussis and to have paroxysmal cough, posttussive vomiting, whooping and a cough duration of > or = 4 weeks than Group 2 or 3 subjects. In contrast Group 2 subjects were more likely than Group 1 subjects to have had previous pertussis immunization or prior antibiotics. PCR identified many mild illnesses caused by B. pertussis that were not identified by culture.
在德国进行的一项大型疫苗效力试验中,聚合酶链反应(PCR)最近被添加到传统培养和血清学检测方法中,用于诊断百日咳博德特氏菌感染。对于出现咳嗽症状的疫苗接种者或家庭成员,采集两份鼻咽拭子(一份用于培养的海藻酸钙拭子和一份用于PCR的涤纶拭子),并获取初始和随访临床数据。使用寡核苷酸引物PTp1和PTp2进行PCR,该引物可扩增百日咳毒素操纵子的一个191碱基对的DNA片段。从1993年12月至1994年5月,共处理了555对拭子;28份培养出百日咳博德特氏菌,9份培养出副百日咳博德特氏菌。28例百日咳博德特氏菌培养阳性的受试者中有26例PCR结果也呈阳性,9例副百日咳博德特氏菌病例中有1例PCR结果阳性,另有82份样本PCR结果阳性。PCR使百日咳博德特氏菌感染受试者的识别率提高了近4倍。根据实验室检测结果类别分析临床特征(第1组,28例培养阳性;第2组,82例培养阴性、PCR阳性;第3组,436例培养和PCR均阴性)。与第2组或第3组受试者相比,第1组受试者更有可能被诊断为确诊或疑似百日咳,更有可能出现阵发性咳嗽、咳嗽后呕吐、哮吼,且咳嗽持续时间≥4周。相比之下,第2组受试者比第1组受试者更有可能既往接种过百日咳疫苗或使用过抗生素。PCR识别出许多由百日咳博德特氏菌引起的轻症病例,而这些病例通过培养无法识别。