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Design and validation of a multiplex PCR protocol for microsatellite typing of Candida parapsilosis sensu stricto isolates

Este es el resumen de nuestro último artículo científico publicado en BMC GenomicsQuiero agradecer a mis colegas de la Universidad del País Vasco / Euskal Herriko Unibertsitatea Camino Trobajo Sanmartín Elena Eraso, a Célia Pais, del Centro de Biologia Molecular e Ambiental (CBMA), Departamento de Biologia, Universidade do Minho, Braga, Portugal, y a Guillermo Ezpeleta del Servicio de Medicina Preventiva e Higiene Hospitalaria, Complejo Hospitalario de Navarra, Pamplona, su inestimable colaboración y coautoría.

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Design and validation of a multiplex PCR protocol for microsatellite typing of Candida parapsilosis sensu stricto isolates. Summary

Background

Analysis of polymorphic microsatellite markers (STR) is a helpful genotyping technique to differentiate Candida parapsilosis sensu stricto isolates. The aim of this study is to develop and perform an initial validation of an alternative protocol for the reliable and accurate microsatellite genotyping of C. parapsilosis sensu stricto isolates using high-throughput multiplex PCR. To achieve this, the results obtained using the new protocol were compared to the ones obtained using a previously described reference method. To that end, diagnostic accuracy, informativeness and discrimination parameters were estimated.

Results

Our results showed good concordance between both methods (Kappa index: 0.920), leading to a high sensitivity (1; CI(95%) (0.991–1)) and specificity (1; CI(95%) (0.772–1)) after the validation of the new protocol. Moreover, the electropherograms profiles obtained with the new PCR scheme showed a high signal to noise ratio (SNR).

Conclusions

The new multiplex protocol is valuable for the differentiation of C. parapsilosis sensu stricto, with direct clinical applications. Besides, the new protocol represents a shortening the hands-on time, reducing the sample manipulation (dismissing the possibility of cross-contamination), maintaining the quality of the results (when compared to the ones obtained with the reference method), and helping to the standardization and simplification of the genotyping scheme.

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