Establishment of Detection Methodology of C.albicans by Real-time PCR

Du Fei; Li Rong; He Rui; Zeng Shui Qin; Duan Kai Wen; Li Cheng Wen

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Du Fei , Li Rong , He Rui , Zeng Shui Qin , Duan Kai Wen , Li Cheng Wen . Establishment of Detection Methodology of C.albicans by Real-time PCR[J]. Journal of Kunming Medical University, 2017, 38(07): 55-59.

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Du Fei , Li Rong , He Rui , Zeng Shui Qin , Duan Kai Wen , Li Cheng Wen . Establishment of Detection Methodology of C.albicans by Real-time PCR[J]. Journal of Kunming Medical University, 2017, 38(07): 55-59.

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Establishment of Detection Methodology of C.albicans by Real-time PCR

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基金：国家自然科学基金资助项目 (30860315, 81160135); 云南省科技厅-昆明医科大学联合专项基金资助项目 (2015FA009);

Received Date: 2017-02-21

Abstract

Abstract

Objective To use TaqMan MGB probe technology to establish a detection methodology of Candida albicans (C.albicans) by real-time fluorescence quantitative PCR (RQ-PCR) and to provide a quick method for the detection of candida infection in clinical settings. Methods D1/D2 region of candida species which is a highly conserved sequence gene was selected as amplification target sequence and specific primers and probes for C.albicans were synthesized. The DNA of standard strains of C.albicans was extracted and rebuilt plasmid containing the target gene fragment was constructed. The DNA was amplified as a standard to establish the standard curve of C.albicans and methodological evaluation was conducted to identify the linear range, repeatability, sensitivity, specificity and so on. Results After sequencing the recombinant cloning vector, the sequence of the fragment inserted was consistent with the expected sequence, with the similarity of 100%. C.albicans standard curve Y =-3.824 x +41.36, R2= 0.990, which was in good shape and illustrated the detection method of C.albicans by real-time PCR had successfully established. Conclusion The detection method of C.albicans by real-time PCR has better sensitivity and specificity.
- Real-time quantitative PCR,
- Candida albicans,
- Methodology

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References(20)

References

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