Validation of High Resolution Melting Analysis (HRM) of the Amplified ITS2 Region for the Detection and Identification of Yeasts from Clinical Samples: Comparison with Culture and MALDI-TOF Based Identification

被引：16

作者：

Duyvejonck, Hans ^{[1
,6
]}

Cools, Piet ^{[1
]}

Decruyenaere, Johan ^{[2
]}

Roelens, Kristien ^{[3
]}

Noens, Lucien ^{[4
,5
]}

Vermeulen, Stefan ^{[6
]}

Claeys, Geert ^{[7
]}

Decat, Ellen ^{[1
]}

Van Mechelen, Els ^{[6
]}

Vaneechoutte, Mario ^{[1
]}

机构：

[1] Univ Ghent, Dept Clin Chem Microbiol & Immunol, Lab Bacteriol Res, Fac Med & Hlth Sci, B-9000 Ghent, Belgium

[2] Ghent Univ Hosp, Dept Intens Care Med, B-9000 Ghent, Belgium

[3] Ghent Univ Hosp, Fac Med & Hlth Sci, Dept Obstet & Gynaecol, B-9000 Ghent, Belgium

[4] Ghent Univ Hosp, Dept Haematol, B-9000 Ghent, Belgium

[5] Ghent Univ Hosp, Blood Bank, B-9000 Ghent, Belgium

[6] Univ Coll Ghent, Fac Educ Hlth & Social Work, Dept Biomed Sci, B-9000 Ghent, Belgium

[7] Ghent Univ Hosp, Dept Clin Chem Microbiol & Immunol, B-9000 Ghent, Belgium

来源：

PLOS ONE | 2015年 / 10卷 / 08期

关键词：

SACCHAROMYCES-CEREVISIAE; CANDIDA; RESISTANCE; INFECTIONS; PREVALENCE; DIAGNOSIS; TIME;

D O I：

10.1371/journal.pone.0132149

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Aim Candida species are known as opportunistic pathogens, and a possible cause of invasive infections. Because of their species-specific antimycotic resistance patterns, reliable techniques for their detection, quantification and identification are needed. We validated a DNA amplification method for direct detection of Candida spp. from clinical samples, namely the ITS2-High Resolution Melting Analysis (direct method), by comparing it with a culture and MALDI-TOF Mass Spectrometry based method (indirect method) to establish the presence of Candida species in three different types of clinical samples. Materials and Methods A total of 347 clinical samples, i.e. throat swabs, rectal swabs and vaginal swabs, were collected from the gynaecology/obstetrics, intensive care and haematology wards at the Ghent University Hospital, Belgium. For the direct method, ITS2-HRM was preceded by Nucli-SENS easyMAG DNA extraction, directly on the clinical samples. For the indirect method, clinical samples were cultured on Candida ID and individual colonies were identified by MALDI-TOF. Results For 83.9% of the samples there was complete concordance between both techniques, i.e. the same Candida species were detected in 31.1% of the samples or no Candida species were detected in 52.8% of the samples. In 16.1% of the clinical samples, discrepant results were obtained, of which only 6.01% were considered as major discrepancies. Discrepancies occurred mostly when overall numbers of Candida cells in the samples were low and/or when multiple species were present in the sample. Discussion Most of the discrepancies could be decided in the advantage of the direct method. This is due to samples in which no yeast could be cultured whereas low amounts could be detected by the direct method and to samples in which high quantities of Candida robusta according to ITS2-HRM were missed by culture on Candida ID agar. It remains to be decided whether the diagnostic advantages of the direct method compensate for its disadvantages.

引用

页数：16