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See detailDirect detection of Aspergillus and azole resistance of Aspergillus fumigatus on broncho-alveolar lavage fluid. Validation of a new Aspergillus real-time PCR
Chong, Ga-Lai; Van de Sande, Wendy; Dingemans, Gijs et al

in Journal of Clinical Microbiology (2015)

Introduction Azole resistance in Aspergillus fumigatus is increasingly reported. We describe the validation of AsperGenius® , a new multiplex real-time polymerase chain reaction (PCR) assay consisting of ... [more ▼]

Introduction Azole resistance in Aspergillus fumigatus is increasingly reported. We describe the validation of AsperGenius® , a new multiplex real-time polymerase chain reaction (PCR) assay consisting of two multiplex real-time PCRs: one which identifies the clinically relevant Aspergillus species, and one which detects the TR34, L98H, T289A, Y121F mutations in CYP51A and differentiates susceptible from resistant A. fumigatus strains. Methods The diagnostic performance was tested on 37 bronchoalveolar lavage (BAL) samples from haematology patients and on 40 BAL samples from intensive care unit (ICU) patients using BAL galactomannan ≥1.0 or positive culture as the gold standard for the presence of Aspergillus. Results In the haematology and ICU groups combined, there were 22 BAL samples with IA (2 proven, 9 probable and 11 non-classifiable). Nineteen of the 22 BAL samples were positive according to the gold standard. The optimal cycle threshold value for the presence of Aspergillus was <36. Sixteen of the 19 BAL samples had a positive PCR (2 Aspergillus species and 14 A. fumigatus). This resulted in a sensitivity, specificity, positive and negative predictive value of 88.9%, 89.3%, 72.7% and 96.2% for the haematology group and 80.0%, 93.3%, 80.0% and 93.3% in the ICU group, respectively. The CYP51A real-time PCR confirmed 12 wildtype and 2 resistant strains (1 TR34/L98H and 1 TR46/Y121F/T289A mutant). Conclusion The AsperGenius® multiplex real-time PCR allows for a sensitive and fast detection of Aspergillus species directly in BAL samples. More importantly, this assay detects and differentiates wildtype from resistant strains even if BAL cultures remained negative. [less ▲]

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See detailDevelopment of a new commercial qPCR assay to detect and differentiate dermatophyte infections of the skin, nails and hair
Dingemans, Gijs; van den Bosch, Mélanie; HAYETTE, Marie-Pierre ULg et al

Poster (2014, May)

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See detailIncrease in viral load, viral integration, and gain of telomerase genes during uterine cervical carcinogenesis can be simultaneously assessed by the HPV 16/18 MLPA-assay.
Theelen, Wendy; Speel, Ernst*-Jan M; Herfs, Michael ULg et al

in American Journal of Pathology (2010), 177(4), 2022-33

Oncogenic human papillomavirus (HPV) infection is the most important risk factor in cervical carcinogenesis cases; high viral loads, viral integration into the host genome, and gain of the telomerase ... [more ▼]

Oncogenic human papillomavirus (HPV) infection is the most important risk factor in cervical carcinogenesis cases; high viral loads, viral integration into the host genome, and gain of the telomerase-related genes, TERT and TERC, are all factors associated with progression to cancer. A recently developed multiparameter HPV 16/18 multiplex ligation-dependent probe amplification (MLPA) assay, which allows the simultaneous assessment of these factors, was applied to a series of 67 normal and (pre)malignant frozen uterine cervical samples, as well as to 91 cytological preparations, to test the ability of the MLPA assay to identify high-risk lesions on the basis of these factors. Validation was performed using quantitative PCR, the PapilloCheck and fluorescence in situ hybridization. Only 5 out of 37 normal tissue samples or low-grade cervical lesions (ie, CIN1 and condyloma) showed either an HPV16 viral load higher than 25 copies per cell, viral integration, and/or gain of one of the telomerase-related genes, whereas for the high-grade cervical lesions, one or more of these risk factors was found in 25 of 30 cases. The HPV MLPA assay showed a sensitivity of 83% and a specificity of 86% in frozen cervical specimens. Furthermore, the feasibility of the MLPA assay was shown for cytological samples, where in 57% of high-grade squamous intraepithelial lesion cases, the high-risk factors were detected using this assay. [less ▲]

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See detailA MLPA-oligochromatographic multiplex assay for identification of the toxin genotype and the hypervirulent ribotype O27 of Clostridium difficile
Durenne, Bastien ULg; Laurent, Thierry; Leclipteux, Thierry et al

in Clinical Microbiology & Infection (2009, May), 15(s4),

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