Spinaci Cinzia, Magi Gloria, Varaldo Pietro E, Facinelli Bruna
Institute of Microbiology and Biomedical Sciences, Marche Polytechnic University Medical School, Ancona, Italy.
Pediatr Infect Dis J. 2006 Oct;25(10):880-3. doi: 10.1097/01.inf.0000238136.63851.4a.
A significant association between erythromycin resistance and ability of bacteria to enter human respiratory cells has been documented in group A streptococci (GAS) isolated in Italy from children with pharyngitis. The occurrence of strains combining cell invasiveness with erythromycin resistance raised serious concern because they could escape penicillin by virtue of intracellular location and macrolides by virtue of resistance, resulting in difficulty in eradication.
Thirty-one pharyngeal cell-invasive, erythromycin-resistant (ER) GAS, representing that many clones recently identified among Italian GAS carrying the internalization-related gene prtF1, were investigated for intracellular persistence inside cultured respiratory cells (A549) by means of a survival assay and by staining and polymerase chain reaction assays on infected cells.
All tested ER GAS could be recovered in culture from infected cells 24 hours from infection with logarithms exceeding 4.0 (4 strains). The highest survival rate (>5.0) was exhibited by strain SP1900 [erm(A)/iMLS-B; high-level erythromycin resistance [minimum inhibitory concentration, > or =128 microg/mL)], the most widespread clone, which was cultivable from infected cells up to day 5. As long as SP1900-infected cells could be maintained in culture, the presence of multiple cocci inside cells was consistently revealed by microscopy. During the same time, DNA sequences internal to both genes prtF1 and erm(A) continued to be amplified by polymerase chain reaction.
These results suggest that the ER GAS are capable of persisting in human respiratory cells. This might contribute to clinical problems such as relapsing infection and persistent carriage despite antibiotic treatment and might have facilitated the widespread diffusion of ER GAS in Italy.
在意大利从咽炎患儿中分离出的A组链球菌(GAS)中,已证明红霉素耐药性与细菌进入人呼吸道细胞的能力之间存在显著关联。兼具细胞侵袭性和红霉素耐药性的菌株的出现引起了严重关注,因为它们可能凭借细胞内定位逃避青霉素的作用,并凭借耐药性逃避大环内酯类药物的作用,从而导致难以根除。
通过存活试验以及对感染细胞进行染色和聚合酶链反应分析,对31株具有咽部细胞侵袭性、红霉素耐药(ER)的GAS进行研究,这些菌株代表了最近在携带内化相关基因prtF1的意大利GAS中鉴定出的许多克隆,以检测它们在培养的呼吸道细胞(A549)内的持续存在情况。
所有测试的ER GAS在感染后24小时均可从感染细胞的培养物中回收,对数超过4.0(4株)。最广泛传播的克隆株SP1900 [erm(A)/iMLS-B;高水平红霉素耐药性(最低抑菌浓度,≥128μg/mL)]表现出最高的存活率(>5.0),可从感染细胞中培养至第5天。只要感染SP1900的细胞能够在培养物中维持,显微镜检查就始终能发现细胞内存在多个球菌。与此同时,prtF1和erm(A)这两个基因内部的DNA序列通过聚合酶链反应持续被扩增。
这些结果表明,ER GAS能够在人呼吸道细胞中持续存在。这可能导致诸如复发性感染和尽管接受抗生素治疗仍持续携带等临床问题,并可能促进了ER GAS在意大利的广泛传播。