沙眼衣原体培养阳性率提高及临床株热休克蛋白60的转录随传代的变化
发布时间：2018-07-13 17:33
【摘要】：沙眼衣原体(Chlamydia trachomatis, Ct)感染是目前发达最常见的性病病原体,可引起尿道炎、宫颈炎、子宫内膜炎、附睾炎、前列腺炎、不孕症多种疾病,且发病率呈逐年上升趋势,已经成为一个危害极大的公共健康问题。沙眼衣原体检测的金标准目前仍然是细胞培养。但是细胞培养并未在临床工作中广泛采用。而且,临床株的培养是体外药物敏感试验的基础。因此,提高沙眼衣原体细胞培养检出的阳性率显得尤为重要。另外,导致沙眼衣原体感染治疗失败的一个重要原因是沙眼衣原体的持续感染,持续感染时碘染不能看到典型的包涵体且沙眼衣原体热休克蛋白60含量增加。 目的：1、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聚乙二醇对沙眼衣原体D-UW-5/Cx型、E-UW-5/Cx型标准株4种常用抗菌药物阿奇霉素、米诺环素、莫西沙星、多西环素体外药物敏感性和31例临床标本包涵体数量的影响。 2、探讨E型标准株和40例经细胞培养成功的沙眼衣原体E型临床株随着传代次数的增加其热休克蛋白60(HSP60)含量的改变及不同治疗效果患者首次传代碘染出现包涵体情况的差别。 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的细胞培养液,离心后在孵箱中静置2h后换成衣原体感染液,48h后固定,碘染计数包涵体数量。将经过0.7%PEG处理过的沙眼衣原体接种于细胞培养板的McCoy细胞,使McCoy细胞感染率达90%以上后进行药敏实验。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并在细胞培养液中加入0.7%PEG,将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细胞后计数第一代全孔包涵体数量。 将经McCoy细胞培养法检测出的40例E型沙眼衣原体临床株,通过RT-PCR方法检测首次传代,二次传代,三次传代和四次传代的CHSP60含量,将40例E型沙眼衣原体临床株分为首次传代出现和未出现包涵体两组,通过χ2检验不同治疗效果患者首次传代碘染出现包涵体情况的差别。 结果：0.7%PEG能使沙眼衣原体E型标准株包涵体数量提高3.44倍,D型标准株包涵体数量提高3.56倍,为促进沙眼衣原体D、E型生长的最适浓度。在体外,PEG处理过的沙眼衣原体D、E型标准株的阿奇霉素、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。经细胞培养成功的E型沙眼衣原体临床株传代未出现包涵体时其CHSP60含量较出现包涵体时高(P0.05)。首次传代出现包涵体与否其对应患者治疗效果有差别,治疗失败患者80%首次传代未出现包涵体,一个疗程治疗后转阴患者70%在首次传代出现包涵体。 结论：1、0.7%的PEG可以显著促进沙眼衣原体D型、E型的生长并对沙眼衣原体D型、E型标准株药敏结果无明显影响,且可以进一步提高临床标本检测的敏感性。 2、80%治疗失败患者首次传代未见包涵体,经过细胞培养成功但首次或者二次传代培养未出现包涵体的临床标本其对应患者感染的衣原体其合成CHSP60增加,二者提示衣原体可能处于一个持续性状态,出现包涵体可能标志着这种持续性状态的解除。
[Abstract]:Chlamydia pneumoniae ( Ct ) infection is the most common pathogen of sexually transmitted diseases , which can cause urinary tract inflammation , cervicitis , endometritis , epimitis , prostatitis and infertility , and has become a major public health problem .
Objective : 1 . To investigate the effects of polyethylene glycol ( PEG ) on the growth of C . trachoma - 5 / Cx - type , E - UWB - 5 / Cx - type standard strains , and the effect of polyethylene glycol on the drug susceptibility and the number of clinical specimens in 31 clinical specimens .
2 . To investigate the changes of heat shock protein 60 ( HSP60 ) and the difference of inclusion bodies in the first passage of iodine staining in E - type standard strains and 40 cases of Chlamydia - Chlamydia E - type clinical strains with successful passaging .
Methods : In the McCoy cells with different concentrations of PEG were added to the McCoy cells with different concentrations of PEG on the D - and E - type strains of Chlamydia trachoma .
40 cases of E - type trachoma chlamydia were detected by the McCoy cell culture method , and 40 cases of E - type trachoma chlamydia were divided into two groups : first passage appearance and no inclusion body , and the difference of inclusion body in the first passage of iodine staining was tested by 蠂2 test .
Results : 0 . 7 % PEG could increase the number of inclusion bodies of the E - type standard strain of Chlamydia trachoma by 3.44 times , and the number of inclusion bodies of D - type standard strains increased by 3.56 times .
Conclusion : 1 銆，
本文编号：2120190
[Abstract]:Chlamydia pneumoniae ( Ct ) infection is the most common pathogen of sexually transmitted diseases , which can cause urinary tract inflammation , cervicitis , endometritis , epimitis , prostatitis and infertility , and has become a major public health problem .
Objective : 1 . To investigate the effects of polyethylene glycol ( PEG ) on the growth of C . trachoma - 5 / Cx - type , E - UWB - 5 / Cx - type standard strains , and the effect of polyethylene glycol on the drug susceptibility and the number of clinical specimens in 31 clinical specimens .
2 . To investigate the changes of heat shock protein 60 ( HSP60 ) and the difference of inclusion bodies in the first passage of iodine staining in E - type standard strains and 40 cases of Chlamydia - Chlamydia E - type clinical strains with successful passaging .
Methods : In the McCoy cells with different concentrations of PEG were added to the McCoy cells with different concentrations of PEG on the D - and E - type strains of Chlamydia trachoma .
40 cases of E - type trachoma chlamydia were detected by the McCoy cell culture method , and 40 cases of E - type trachoma chlamydia were divided into two groups : first passage appearance and no inclusion body , and the difference of inclusion body in the first passage of iodine staining was tested by 蠂2 test .
Results : 0 . 7 % PEG could increase the number of inclusion bodies of the E - type standard strain of Chlamydia trachoma by 3.44 times , and the number of inclusion bodies of D - type standard strains increased by 3.56 times .
Conclusion : 1 銆，
本文编号：2120190
本文链接：https://www.wllwen.com/yixuelunwen/pifb/2120190.html
最近更新
教材专著
