中国实验诊断学
中國實驗診斷學
중국실험진단학
CHINESE JOURNAL OF LABORATORY DIAGNOSIS
2015年
4期
528-531
,共4页
许凡凡%贺丹%孙晓红%王丽%高嵩%盛辉
許凡凡%賀丹%孫曉紅%王麗%高嵩%盛輝
허범범%하단%손효홍%왕려%고숭%성휘
白假丝酵母%荧光定量 PCR%鉴定%COⅡ基因
白假絲酵母%熒光定量 PCR%鑒定%COⅡ基因
백가사효모%형광정량 PCR%감정%COⅡ기인
Candida albicans%real-time PCR%identification%COⅡ gene
目的：真菌感染发病率和死亡率逐年增高，亟需建立主要病原真菌快速、敏感、特异的鉴定方法，对感染的早期诊治至关重要。方法以真菌线粒体 COⅡ基因为靶标，设计白假丝酵母的特异性引物，建立荧光定量 PCR 鉴定方法，分析其灵敏度、特异性及重复性。结果该方法检测的灵敏度可达1×102 copies／μl（相当于10－7 ng／μl），熔解曲线显示具有单一峰，且仅有白假丝酵母出现扩增曲线，其他种、属真菌均未见扩增。且具有较好的重复性和稳定性。结论建立的白假丝酵母荧光定量 PCR 鉴定方法灵敏度高、特异性强、重复性和稳定性好，适于临床真菌感染微量标本的快速检测。该研究为真菌核酸快速检测试剂盒的开发奠定了基础。
目的：真菌感染髮病率和死亡率逐年增高，亟需建立主要病原真菌快速、敏感、特異的鑒定方法，對感染的早期診治至關重要。方法以真菌線粒體 COⅡ基因為靶標，設計白假絲酵母的特異性引物，建立熒光定量 PCR 鑒定方法，分析其靈敏度、特異性及重複性。結果該方法檢測的靈敏度可達1×102 copies／μl（相噹于10－7 ng／μl），鎔解麯線顯示具有單一峰，且僅有白假絲酵母齣現擴增麯線，其他種、屬真菌均未見擴增。且具有較好的重複性和穩定性。結論建立的白假絲酵母熒光定量 PCR 鑒定方法靈敏度高、特異性彊、重複性和穩定性好，適于臨床真菌感染微量標本的快速檢測。該研究為真菌覈痠快速檢測試劑盒的開髮奠定瞭基礎。
목적：진균감염발병솔화사망솔축년증고，극수건립주요병원진균쾌속、민감、특이적감정방법，대감염적조기진치지관중요。방법이진균선립체 COⅡ기인위파표，설계백가사효모적특이성인물，건립형광정량 PCR 감정방법，분석기령민도、특이성급중복성。결과해방법검측적령민도가체1×102 copies／μl（상당우10－7 ng／μl），용해곡선현시구유단일봉，차부유백가사효모출현확증곡선，기타충、속진균균미견확증。차구유교호적중복성화은정성。결론건립적백가사효모형광정량 PCR 감정방법령민도고、특이성강、중복성화은정성호，괄우림상진균감염미량표본적쾌속검측。해연구위진균핵산쾌속검측시제합적개발전정료기출。
Objective The morbidity and mortality of fungal infections increased these years.It is necessary to es-tablish a rapid,sensitive and specific identification method which is essential for early diagnosis and treatment of the in-fections.Methods Specific primers of Candida albicans were designed according to the sequences of fungal mitochon-drial COⅡ gene.And a real-time PCR method was established to identify Candida albicans .The sensitivity,specificity and repeatability of this identification method were analyzed.Results Sensitivity of the method was up to 1 × 102 copies/μl (equivalent 10-7 ng/μl).There was a single peak of dissociation curve.Only Candida albicans was ampli-fied,other genus and speciesof fungi were not amplified.The method had good repeatability and stability.Conclusion The established real-time PCR method of Candida albicans has higher sensitivity and specificity,good repeatability and stability,which is suitable for rapid diagnosis with trace specimens of fungal infections in clinic.This study will lay the foundation for the development of rapid diagnostic kit of fungi.
