Unité de Biologie des Spirochètes, Institut Pasteur, Paris, France.
PLoS Negl Trop Dis. 2010 Jun 22;4(6):e724. doi: 10.1371/journal.pntd.0000724.
Leptospirosis has been implicated as a severe and fatal form of disease in Mayotte, a French-administrated territory located in the Comoros archipelago (southwestern Indian Ocean). To date, Leptospira isolates have never been isolated in this endemic region.
Leptospires were isolated from blood samples from 22 patients with febrile illness during a 17-month period after a PCR-based screening test was positive. Strains were typed using hyper-immune antisera raised against the major Leptospira serogroups: 20 of 22 clinical isolates were assigned to serogroup Mini; the other two strains belonged to serogroups Grippotyphosa and Pyrogenes, respectively. These isolates were further characterized using partial sequencing of 16S rRNA and ligB gene, Multi Locus VNTR Analysis (MLVA), and pulsed field gel electrophoresis (PFGE). Of the 22 isolates, 14 were L. borgpetersenii strains, 7 L. kirschneri strains, and 1, belonging to serogoup Pyrogenes, was L. interrogans. Results of the genotyping methods were consistent. MLVA defined five genotypes, whereas PFGE allowed the recognition of additional subgroups within the genotypes. PFGE fingerprint patterns of clinical strains did not match any of the patterns in the reference strains belonging to the same serogroup, suggesting that the strains were novel serovars.
Preliminary PCR screening of blood specimen allowed a high isolation frequency of leptospires among patients with febrile illness. Typing of leptospiral isolates showed that causative agents of leptospirosis in Mayotte have unique molecular features.
在法属马约特岛(位于印度洋西南部科摩罗群岛),钩端螺旋体病被认为是一种严重且致命的疾病形式。迄今为止,在这个地方性流行地区从未分离出钩端螺旋体。
在基于 PCR 的筛查试验阳性后的 17 个月期间,从 22 名发热患者的血液样本中分离出了钩端螺旋体。使用针对主要钩端螺旋体血清群的高免疫抗血清对菌株进行分型:22 株临床分离株中有 20 株被分配到血清群 Mini;另外两株分别属于血清群 Grippotyphosa 和 Pyrogenes。使用 16S rRNA 和 ligB 基因的部分测序、多位点可变数目串联重复分析(MLVA)和脉冲场凝胶电泳(PFGE)进一步对这些分离株进行了特征描述。在 22 株分离株中,有 14 株为 L. borgpetersenii 菌株,7 株为 L. kirschneri 菌株,1 株属于血清群 Pyrogenes,为 L. interrogans。基因分型方法的结果一致。MLVA 定义了五个基因型,而 PFGE 允许在同一基因型内识别额外的亚群。临床菌株的 PFGE 指纹图谱与属于同一血清群的参考菌株的任何图谱均不匹配,表明这些菌株是新型血清型。
对血液标本进行初步 PCR 筛查可大大提高发热患者中钩端螺旋体的分离率。钩端螺旋体分离株的分型表明,马约特岛钩端螺旋体病的病原体具有独特的分子特征。