【摘要】：沙眼衣原體(Chlamydia trachomatis, Ct)感染是目前發(fā)達(dá)最常見的性病病原體,可引起尿道炎、宮頸炎、子宮內(nèi)膜炎、附睪炎、前列腺炎、不孕癥多種疾病,且發(fā)病率呈逐年上升趨勢,已經(jīng)成為一個危害極大的公共健康問題。沙眼衣原體檢測的金標(biāo)準(zhǔn)目前仍然是細(xì)胞培養(yǎng)。但是細(xì)胞培養(yǎng)并未在臨床工作中廣泛采用。而且,臨床株的培養(yǎng)是體外藥物敏感試驗(yàn)的基礎(chǔ)。因此,提高沙眼衣原體細(xì)胞培養(yǎng)檢出的陽性率顯得尤為重要。另外,導(dǎo)致沙眼衣原體感染治療失敗的一個重要原因是沙眼衣原體的持續(xù)感染,持續(xù)感染時碘染不能看到典型的包涵體且沙眼衣原體熱休克蛋白60含量增加。 目的：1、探討聚乙二醇(PEG)促進(jìn)沙眼衣原體D-UW-5/Cx型、E-UW-5/Cx型標(biāo)準(zhǔn)株生長的最適濃度及聚乙二醇對沙眼衣原體D-UW-5/Cx型、E-UW-5/Cx型標(biāo)準(zhǔn)株4種常用抗菌藥物阿奇霉素、米諾環(huán)素、莫西沙星、多西環(huán)素體外藥物敏感性和31例臨床標(biāo)本包涵體數(shù)量的影響。 2、探討E型標(biāo)準(zhǔn)株和40例經(jīng)細(xì)胞培養(yǎng)成功的沙眼衣原體E型臨床株隨著傳代次數(shù)的增加其熱休克蛋白60(HSP60)含量的改變及不同治療效果患者首次傳代碘染出現(xiàn)包涵體情況的差別。 方法：在沙眼衣原體D、E型菌株接種于致密單層的McCoy細(xì)胞時,加入含有不同濃度PEG的細(xì)胞培養(yǎng)液,離心后在孵箱中靜置2h后換成衣原體感染液,48h后固定,碘染計數(shù)包涵體數(shù)量。將經(jīng)過0.7%PEG處理過的沙眼衣原體接種于細(xì)胞培養(yǎng)板的McCoy細(xì)胞,使McCoy細(xì)胞感染率達(dá)90%以上后進(jìn)行藥敏實(shí)驗(yàn)。采用微量稀釋法測定體外4種抗菌藥物對沙眼衣原體的作用并在細(xì)胞培養(yǎng)液中加入0.7%PEG,將31例衣原體細(xì)胞培養(yǎng)陽性且基因分型為D、E型的臨床標(biāo)本加入或者不加入0.7%PEG接種于致密單層的McCoy細(xì)胞后計數(shù)第一代全孔包涵體數(shù)量。 將經(jīng)McCoy細(xì)胞培養(yǎng)法檢測出的40例E型沙眼衣原體臨床株,通過RT-PCR方法檢測首次傳代,二次傳代,三次傳代和四次傳代的CHSP60含量,將40例E型沙眼衣原體臨床株分為首次傳代出現(xiàn)和未出現(xiàn)包涵體兩組,通過χ2檢驗(yàn)不同治療效果患者首次傳代碘染出現(xiàn)包涵體情況的差別。 結(jié)果：0.7%PEG能使沙眼衣原體E型標(biāo)準(zhǔn)株包涵體數(shù)量提高3.44倍,D型標(biāo)準(zhǔn)株包涵體數(shù)量提高3.56倍,為促進(jìn)沙眼衣原體D、E型生長的最適濃度。在體外,PEG處理過的沙眼衣原體D、E型標(biāo)準(zhǔn)株的阿奇霉素、莫西沙星、多西環(huán)素、米諾環(huán)素的最低抑菌濃度與未經(jīng)PEG處理的沙眼衣原體標(biāo)準(zhǔn)株藥敏結(jié)果一致,0.7%PEG可以顯著增加沙眼衣原體D、E型臨床標(biāo)本傳代產(chǎn)生的包涵體數(shù)量。經(jīng)細(xì)胞培養(yǎng)成功的E型沙眼衣原體臨床株傳代未出現(xiàn)包涵體時其CHSP60含量較出現(xiàn)包涵體時高(P0.05)。首次傳代出現(xiàn)包涵體與否其對應(yīng)患者治療效果有差別,治療失敗患者80%首次傳代未出現(xiàn)包涵體,一個療程治療后轉(zhuǎn)陰患者70%在首次傳代出現(xiàn)包涵體。 結(jié)論：1、0.7%的PEG可以顯著促進(jìn)沙眼衣原體D型、E型的生長并對沙眼衣原體D型、E型標(biāo)準(zhǔn)株藥敏結(jié)果無明顯影響,且可以進(jìn)一步提高臨床標(biāo)本檢測的敏感性。 2、80%治療失敗患者首次傳代未見包涵體,經(jīng)過細(xì)胞培養(yǎng)成功但首次或者二次傳代培養(yǎng)未出現(xiàn)包涵體的臨床標(biāo)本其對應(yīng)患者感染的衣原體其合成CHSP60增加,二者提示衣原體可能處于一個持續(xù)性狀態(tài),出現(xiàn)包涵體可能標(biāo)志著這種持續(xù)性狀態(tài)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，

本文編號：2120190