Stefanelli P, De Marzi L, Robino L, Roman D, Basso F, D'Orazio P, Di Tommaso S, Vasile E, Caciolo E, Spigaglia P, Mastrantonio P
Laboratory of Bacteriology and Medical Mycology, Istituto Superiore di Sanità, Rome, Italy.
New Microbiol. 1999 Jul;22(3):187-94.
During the clinical trial conducted in Italy to evaluate the efficacy of new acellular pertussis vaccines, the most favorable conditions for the recovery and characterization of Bordetella pertussis strains, isolated from children with cough, were adopted. The nasopharyngeal aspirates were collected and sent to the laboratory in refrigerated conditions within 24 hours. Charcoal agar selective and non selective plates were used, and most of the isolates were recovered after 3-4 days of incubation. Confirmation of all suspected colonies included the use of biochemical tests and specific agglutination reaction with B. pertussis antiserum. Serotyping of fimbriae, susceptibility to erythromycin and DNA fingerprinting by Pulsed Field Gel Electrophoresis (PFGE), were performed to characterize B. pertussis isolates and to determine relatedness among different strains. Serotype 1,3 was the most represented in the bacterial population examined. A predominant pulsetype (PTA) characterized most of the isolates accounting for 71.4% of the strains examined. Eight subclones (23.5%) and three unrelated pulsetypes were also found. No resistant strains to erythromycin were detected.
在意大利进行的一项评估新型无细胞百日咳疫苗疗效的临床试验中,采用了最有利于从咳嗽儿童中分离出的百日咳博德特氏菌菌株进行回收和鉴定的条件。收集鼻咽抽吸物,并在24小时内冷藏送至实验室。使用了木炭琼脂选择性和非选择性平板,大多数分离株在培养3-4天后回收。对所有疑似菌落的确认包括使用生化试验和与百日咳博德特氏菌抗血清的特异性凝集反应。进行菌毛血清分型、对红霉素的敏感性检测以及通过脉冲场凝胶电泳(PFGE)进行DNA指纹图谱分析,以鉴定百日咳博德特氏菌分离株并确定不同菌株之间的相关性。血清型1,3在所检测的细菌群体中最为常见。一种主要的脉冲型(PTA)特征存在于大多数分离株中,占所检测菌株的71.4%。还发现了八个亚克隆(23.5%)和三种不相关的脉冲型。未检测到对红霉素耐药的菌株。