CAJ | 학술논문

目的探讨聚乙二醇(PEG)促进沙眼衣原体D-UW-5/Cx型、E-UW-5/Cx型标准株生长的最适浓度及对4种常用抗菌药物体外药物敏感性的影响.方法在沙眼衣原体D、E型菌株接种于致密单层的McCoy细胞时,加入含有不同浓度PEG的离心液,离心后在孵箱中静置2h后换成衣原体感染液,48 h后固定,碘染计数包涵体数量.将沙眼衣原体接种于McCoy细胞,接种过程中用0.7％(7 g/L)PEG处理菌株,McCoy细胞感染率达90％以上确定沙眼衣原体接种量后进行药敏实验.采用微量稀释法测定体外4种抗菌药物对沙眼衣原体的作用.将取自31例衣原体细胞培养阳性且基因分型为D、E型的临床标本加入或者不加入0.7％PEG,接种于致密单层的McCoy细胞后计数第1代全孔包涵体数量.结果 0.7％PEG能使沙眼衣原体E型包涵体数量提高3.44倍,D型包涵体数量提高3.56倍.在体外,PEG处理过的沙眼衣原体D、E型标准株对阿奇霉素、莫西沙星、多西环素、米诺环素的最低抑菌浓度与未经PEG处理的沙眼衣原体标准株药敏结果一致.0.7％PEG可以显著增加沙眼衣原体D、E型临床标本传代产生的包涵体数量.结论 0.7％PEG可以显著促进沙眼衣原体D型、E型的生长,但对药敏结果无明显影响.
목적 탐토취을이순(PEG)촉진사안의원체D-UW-5/Cx형、E-UW-5/Cx형표준주생장적최괄농도급대4충상용항균약물체외약물민감성적영향.방법 재사안의원체D、E형균주접충우치밀단층적McCoy세포시,가입함유불동농도PEG적리심액,리심후재부상중정치2h후환성의원체감염액,48 h후고정,전염계수포함체수량.장사안의원체접충우McCoy세포,접충과정중용0.7％(7 g/L)PEG처리균주,McCoy세포감염솔체90％이상학정사안의원체접충량후진행약민실험.채용미량희석법측정체외4충항균약물대사안의원체적작용.장취자31례의원체세포배양양성차기인분형위D、E형적림상표본가입혹자불가입0.7％PEG,접충우치밀단층적McCoy세포후계수제1대전공포함체수량.결과 0.7％PEG능사사안의원체E형포함체수량제고3.44배,D형포함체수량제고3.56배.재체외,PEG처리과적사안의원체D、E형표준주대아기매소、막서사성、다서배소、미낙배소적최저억균농도여미경PEG처리적사안의원체표준주약민결과일치.0.7％PEG가이현저증가사안의원체D、E형림상표본전대산생적포함체수량.결론 0.7％PEG가이현저촉진사안의원체D형、E형적생장,단대약민결과무명현영향.
Objective To optimize the concentration of polyethylene glycol (PEG) for the growth of Chlamydia trachomatis (Ct) reference strains D-UW-5/Cx and E-UW-5/Cx,and to evaluate the effects of PEG on the sensitivity of Ct to 4 common antibiotics.Methods After the inoculation of Ct standard strains (D-UW-5/Cx and E-UW-5/Cx) into McCoy cell monolayer,different concentrations of PEG were added into the culture medium followed by centrifugation.After 2 hours of incubation at 37 ℃,the inoculum was removed and a complete culture medium containing 1 μg/mL cycloheximide was added followed by another 48-hour culture.Subsequently,the culture was fixed and subjected to iodine staining for the calculation of Ct inclusions and optimization of PEG for the growth of Ct.Some Ct standard strains were used to infect McCoy cells,with PEG (0.7％,wt/vol) added to the culture medium after inoculation and before centrifugation process,and when the infection rate reached higher than 90％,a microdilution method was utilized to evaluate the minimal inhibitory concentration (MIC) of 4 antibiotics,including azithromycin,minocyline,moxifloxacin and doxycycline.Thirty-one clinical specimens,which had been confirmed to be positive for Ct serovar D or E strain,were inoculated into McCoy cell monolayer for the passage of Ct with or without the presence of 0.7％ PEG.Results The optimal concentration of PEG was 0.7％ for the growth of Ct,and this concentration of PEG could increase the number of inclusion bodies of Ct serovar E by 3.44 folds,and that of Ct serovar D by 3.56 folds.In vitro,the MICs of the 4 antibiotics were consistent between PEG-treated and untreated Ct reference strains.Moreover,PEG notably increased the quantity of inclusion bodies of Ct serovar E or D from clinical specimens after passages.Conclusions PEG (0.7％) can enhance the growth of Ct serovar D and E,but has no obvious influence on antimicrobial susceptibility of Ct.