论文摘要
白假丝酵母菌(Candida albicans),是人体正常菌群之一,在口腔、皮肤、肠道、阴道、肛门等部位均可分离到,是引起人体真菌感染的主要条件致病菌。随着医学科学的发展,各种新药、新技术如广谱抗生素、激素、各种导管插管技术等的广泛使用,导致深部真菌病的发病率愈来愈高,且以C.albicans感染为主。因此,对C.albicans感染的诊断和分型在临床和流行病学研究中都具有重要意义。对C.albicans进行基因分型的目的在于:(1)可追踪传染源,明确传播途径,防止暴发流行;(2)确定同一患者反复感染是源于复发或是再感染;(3)明确某一菌株是否与某种临床表现或疾病相关,从而发现其独特的致病机制;(4)了解感染的发生除与宿主全身或局部易感因素有关外,是否与菌本身的遗传因素有关;(5)了解C.albicans耐药株和敏感株之间基因型的差异、表型和基因型之间的关系等。本研究采用灵敏特异的PCR分型方法,扩增C.albicans的25S rDNA基因的内含子区可转座Ⅰ型内含子插入片段,根据扩增产物的大小和片段数目来分型。A型为野生型C.albicans,无内含子;B型和C型为有379bp转座子的菌株;E型为具有379bp、252bp、341bp三种转座子的菌株,D型为都柏林假丝酵母菌(C.dubliniensis)。药敏实验采用ATB FUNGUS 3酵母菌药物敏感性检测板条,对两性霉素B(Amphotericin B,AMB)、氟康唑(Fluconazol,FCA)、伊曲康唑(Itraconazole,ITR)、5-氟胞嘧啶(5-flurocytosine,5-FC)和伏立康唑(Voriconazole,VRC)5种抗真菌药物进行药敏分析。本研究收集了临床79名患者的痰、咽拭子、大便、血液、尿液、伤口分泌物、脑脊液、静脉插管、穿刺液等标本,从中共分离出C.albicans 90株,对其进行了PCR基因分型鉴定,结果共检出A、B、C三种基因型,未见D、E型,其中A型44株(44/90,48.9%),B型33株(33/90,36.7%),C型13株(13/90,14.4%)。药敏实验结果表明C.albicans的基因型与其抗真菌药物的敏感性之间有一定相关性。各基因型C.albicans均未出现AMB耐药株,两性霉素B仍是治疗危重患者酵母菌感染的首选药;A型对5-FC的敏感率为95.5%,有4.5%耐药;A、B、C三型对FCA的敏感率分别为93.2%、90.9%和76.9%,其中C型C.albicans对FCA的耐药率有高于A型和B型的趋势;A、B、C三型对ITR的敏感率分别为90.8%、81.8%和76.9%,B型和C型C.albicans对ITR的耐药率有高于A型的趋势;A、B、C三型对VRC的敏感率分别为97.7%、96.9%和92.3%,C型C.albicans对VRC的耐药率也有高于A型和B型的趋势。C.albicans PCR分型方法简便、快速、重复性好、特异性强;不同型别的C.albicans耐药谱有差异。弄清C.albicans的基因型与耐药性之间的关系,对Calbicans感染的诊断、耐药性变异、监控医院内真菌感染情况、真菌流行病学研究、指导临床医师合理用药等,具有重要意义。
论文目录
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