In Vivo Expression of Candida Drug Resistance Genes

描述（由申请人提供）： 致病性酵母菌白色念珠菌导致衰弱和经常复发的真菌感染，特别是在免疫受损的患者群体，包括艾滋病患者中。这些感染通常用唑类抗真菌药物治疗，耐药性是一个重要问题，特别是在HIV感染者中。唑类耐药的主要机制包括靶酶的点突变和过表达以及外排泵的过表达。然而，所有关于抗真菌药物耐药性的信息都是从临床分离株和实验室培养物中获得的。这些耐药机制尚未在直接取自患者的样本中得到鉴定或表征。本研究的总体目标是描述C.白念珠菌耐药基因在艾滋病患者的口腔样本，以了解在体内的耐药机制。具体目标是：1。采用定量RT-PCR测定体外培养和体内患者唾液样本中耐药基因的表达水平。2.确定HIV +患者含有敏感和耐药Ca的唾液样本中与耐药相关基因的表达水平。唑类药物与真菌细胞之间的相互作用以及唑类药物的耐药性在可预见的未来仍将是临床上重要的问题。该提案将确定直接从患者样本中的人类样本中采集的敏感和耐药分离株中耐药基因的表达水平，扩展在酵母培养物中进行的观察。这些分析将进一步了解抗真菌药物和靶细胞在宿主环境中的相互作用，并可能导致真菌感染和耐药性的诊断，治疗和预防的改进。

DESCRIPTION (provided by applicant): The pathogenic yeast Candida albicans causes debilitating and often recurring fungal infections, particularly in immune-compromised patient populations including AIDS patients. These infections are usually treated with azole antifungal drugs, and resistance is significant problem, especially in HIV-infected individuals. The major mechanisms of azole resistance include point mutations and overexpression of the target enzyme and overexpression of efflux pumps. However, all of the information concerning antifungal drug resistance has been obtained from studies of clinical isolates and laboratory constructs in culture. These resistance mechanisms have not been identified or characterized in samples taken directly from the patient. The Overall Goal of this research is to characterize the quantitative expression patterns of C. albicans resistance genes in oral samples from HIV patients in order to understand the mechanisms of resistance in vivo. The Specific Aims are: 1. To adapt quantitative RT-PCR to determine levels of expression of resistance genes in vitro in culture and in vivo in patient saliva samples. 2. To determine the levels of expression for genes associated with resistance in saliva samples containing susceptible and resistant Ca from HIV + patients. The interactions between azoles and fungal cells, and azole drug resistance, will continue to be clinically significant issues for the foreseeable future. This proposal will determine the expression levels of resistance genes in susceptible and resistant isolates taken directly from human samples in patient samples, extending the observations made in yeast cultures. These analyses will further the understanding of the interactions of antifungal drugs and the target cells in the context of the host, and may lead to improvements in diagnosis, treatment and prevention of fungal infections and resistance.