Department of Bacteriology and Mycology, Institute of Tropical Medicine Pedro Kourí, Havana, Cuba.
J Clin Microbiol. 2014 Jan;52(1):45-51. doi: 10.1128/JCM.02381-13. Epub 2013 Oct 16.
This study describes the prevalence and genotype distribution of Pneumocystis jirovecii obtained from nasopharyngeal (NP) swabs from immunocompetent Cuban infants and toddlers with whooping cough (WC). A total of 163 NP swabs from 163 young Cuban children with WC who were admitted to the respiratory care units at two pediatric centers were studied. The prevalence of the organism was determined by a quantitative PCR (qPCR) assay targeting the P. jirovecii mitochondrial large subunit (mtLSU) rRNA gene. Genotypes were identified by direct sequencing of mtLSU ribosomal DNA (rDNA) and restriction fragment length polymorphism (RFLP) analysis of the dihydropteroate synthase (DHPS) gene amplicons. qPCR detected P. jirovecii DNA in 48/163 (29.4%) samples. mtLSU rDNA sequence analysis revealed the presence of three different genotypes in the population. Genotype 2 was most common (48%), followed in prevalence by genotypes 1 (23%) and 3 (19%); mixed-genotype infections were seen in 10% of the cases. RFLP analysis of DHPS PCR products revealed four genotypes, 18% of which were associated with resistance to sulfa drugs. Only contact with coughers (prevalence ratio [PR], 3.51 [95% confidence interval {CI}, 1.79 to 6.87]; P = 0.000) and exposure to tobacco smoke (PR, 1.82 [95% CI, 1.14 to 2.92]; P = 0.009) were statistically associated with being colonized by P. jirovecii. The prevalence of P. jirovecii in infants and toddlers with WC and the genotyping results provide evidence that this population represents a potential reservoir and transmission source of P. jirovecii.
本研究描述了从免疫功能正常的古巴婴幼儿百日咳患者的鼻咽(NP)拭子中获得的肺孢子菌(Pneumocystis jirovecii)的流行率和基因型分布。对来自两个儿科中心呼吸护理病房的 163 例患有百日咳的年轻古巴儿童的 163 份 NP 拭子进行了研究。通过针对 P. jirovecii 线粒体大亚基(mtLSU)rRNA 基因的定量 PCR(qPCR)检测确定生物体的流行率。通过 mtLSU 核糖体 DNA(rDNA)的直接测序和二氢叶酸合成酶(DHPS)基因扩增子的限制性片段长度多态性(RFLP)分析来鉴定基因型。qPCR 在 48/163(29.4%)样本中检测到 P. jirovecii DNA。mtLSU rDNA 序列分析显示该人群存在三种不同的基因型。基因型 2 最为常见(48%),其次是基因型 1(23%)和 3(19%);10%的病例存在混合基因型感染。DHPS PCR 产物的 RFLP 分析显示存在四种基因型,其中 18%与对磺胺类药物的耐药性相关。仅与咳嗽者接触(流行率比 [PR],3.51 [95%置信区间 {CI},1.79 至 6.87];P = 0.000)和接触烟草烟雾(PR,1.82 [95% CI,1.14 至 2.92];P = 0.009)与被 P. jirovecii 定植具有统计学相关性。患有百日咳的婴幼儿中 P. jirovecii 的流行率和基因分型结果表明,该人群代表了 P. jirovecii 的潜在储存库和传播源。