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首页> 外文期刊>International Journal of Research in Medical Sciences >Multiplex polymerase chain reaction identification of Candida species colonized sputum of patients suffering from various respiratory tract disorders in Duhok, Iraq
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Multiplex polymerase chain reaction identification of Candida species colonized sputum of patients suffering from various respiratory tract disorders in Duhok, Iraq

机译：伊拉克杜霍克患有各种呼吸道疾病的患者念珠菌物种定殖痰中的多重聚合酶链反应鉴定

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Background: Candida species are part of the body normal flora. Under certain conditions, these opportunistic microorganisms may lead to infection. The purpose of this study was to identify Candida species isolated from sputum from patients suffering from respiratory tract disorders. Methods: A total of 59 sputum samples taken from patients attending Azadi hospital at Duhok province, Kurdistan Region/Iraq. For primary isolation, sputum samples were cultured on sabouraud dextrose agar (SDA). Suspected colonies of Candida isolates were then sub cultured on chromogenic Candida agar for presumptive identification. Genomic DNA extraction was performed using a genomic DNA extraction kit. For rapid identification of Candida spp, specific primers based on the genomic sequence of DNA topoisomerase 11 of C. albicans, C. parapsilosis I, C. parapsilosis II, C. guilliermondi, C. dubliniensis, C. krusei, C. kefyr and C. glabrata, C. tropicalis I, C. tropicalis II, C. lusitaniae were used. The Multiplex PCR products were separated by electrophoresis in 1.5% agarose gel, visualized by staining with ethidium bromide, and photographed. Results: Three Candida species namely C. albicans, C. glabrata and C. tropicalis were differentiated by their colour produced on Chromogenic Candida agar. PCR with the primer mixes yielded 4 different sized of PCR products corresponding to C. albicans, C. glabrata, C. Keyfer and C. tropicalis II, C. glabrata was the most common species (33.33%), followed by C. albicans (16.66%). The highest rate of isolation of Candida species was between the ages of 36 to 45. Conclusion: This study concluded that phenotypic characteristics on selective agar medium such as chromogenic Candida agar are useful for presumptive identification of Candiada spp with the support of molecular method such as multiplex PCR.
机译：背景：念珠菌属人体正常菌群的一部分。在某些条件下，这些机会微生物可能导致感染。这项研究的目的是确定从呼吸道疾病患者的痰中分离出的念珠菌。方法：从伊拉克库克斯坦地区杜霍克省阿扎迪医院就诊的患者中收集的59份痰标本。为了进行初步分离，将痰标本培养在sabouraud葡萄糖琼脂（SDA）上。然后将疑似假丝酵母菌落在发色假丝酵母琼脂上传代培养以进行鉴定。使用基因组DNA提取试剂盒进行基因组DNA提取。为了快速鉴定假丝酵母，基于白色念珠菌，C。parapsilosis I，C。parapsilosis II，C。guilliermondi，C。dubliniensis，C。krusei，C。kefyr和C.使用了glabrata，热带假丝酵母I，热带假丝酵母II，lusitaniae。通过1.5％琼脂糖凝胶电泳分离Multiplex PCR产物，用溴化乙锭染色观察并拍照。结果：通过在产色念珠菌琼脂上产生的颜色区分了白色念珠菌，白色念珠菌和热带念珠菌三种念珠菌。使用引物混合物进行PCR可获得4种不同大小的PCR产物，分别对应于白色念珠菌，光滑念珠菌，Keyfer和C.tropicis II，光滑念珠菌是最常见的物种（33.33％），其次是白色念珠菌（ 16.66％）。念珠菌属物种的最高分离率是在36至45岁之间。结论：本研究得出结论，在选择性琼脂培养基（例如生色念珠菌琼脂）上的表型特征可在分子方法（如分子标记法）的支持下用于假想念珠菌的鉴定。多重PCR。

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