Design and validation of a multiplex PCR protocol for microsatellite typing of Candida parapsilosis sensu stricto isolates
Abstract 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 micr...
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| Format: | Article |
| Language: | English |
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BMC
2018-09-01
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| Series: | BMC Genomics |
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| Online Access: | http://link.springer.com/article/10.1186/s12864-018-5065-3 |
| _version_ | 1818034289216847872 |
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| author | Camino Trobajo-Sanmartín Guillermo Ezpeleta Célia Pais Elena Eraso Guillermo Quindós |
| author_facet | Camino Trobajo-Sanmartín Guillermo Ezpeleta Célia Pais Elena Eraso Guillermo Quindós |
| author_sort | Camino Trobajo-Sanmartín |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-10T06:36:47Z |
| format | Article |
| id | doaj.art-aba321234a2c492180f1bc323251d890 |
| institution | Directory Open Access Journal |
| issn | 1471-2164 |
| language | English |
| last_indexed | 2024-12-10T06:36:47Z |
| publishDate | 2018-09-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Genomics |
| spelling | doaj.art-aba321234a2c492180f1bc323251d8902022-12-22T01:58:54ZengBMCBMC Genomics1471-21642018-09-0119111010.1186/s12864-018-5065-3Design and validation of a multiplex PCR protocol for microsatellite typing of Candida parapsilosis sensu stricto isolatesCamino Trobajo-Sanmartín0Guillermo Ezpeleta1Célia Pais2Elena Eraso3Guillermo Quindós4Laboratorio de Micología Médica, Departamento de Inmunología, Microbiología y Parasitología, UFI11/25 (Microbios y Salud), Facultad de Medicina y Enfermería, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU)Departamento de Medicina Preventiva y Salud Pública, Facultad de Medicina y Enfermería, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU)Centro de Biologia Molecular e Ambiental (CBMA), Departamento de Biologia, Universidade do MinhoLaboratorio de Micología Médica, Departamento de Inmunología, Microbiología y Parasitología, UFI11/25 (Microbios y Salud), Facultad de Medicina y Enfermería, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU)Laboratorio de Micología Médica, Departamento de Inmunología, Microbiología y Parasitología, UFI11/25 (Microbios y Salud), Facultad de Medicina y Enfermería, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU)Abstract 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.http://link.springer.com/article/10.1186/s12864-018-5065-3Candida parapsilosis sensu strictoGenotyping techniqueMicrosatellites repeatsMultiplex PCRSpecificityReproducibility of results |
| spellingShingle | Camino Trobajo-Sanmartín Guillermo Ezpeleta Célia Pais Elena Eraso Guillermo Quindós Design and validation of a multiplex PCR protocol for microsatellite typing of Candida parapsilosis sensu stricto isolates BMC Genomics Candida parapsilosis sensu stricto Genotyping technique Microsatellites repeats Multiplex PCR Specificity Reproducibility of results |
| title | Design and validation of a multiplex PCR protocol for microsatellite typing of Candida parapsilosis sensu stricto isolates |
| title_full | Design and validation of a multiplex PCR protocol for microsatellite typing of Candida parapsilosis sensu stricto isolates |
| title_fullStr | Design and validation of a multiplex PCR protocol for microsatellite typing of Candida parapsilosis sensu stricto isolates |
| title_full_unstemmed | Design and validation of a multiplex PCR protocol for microsatellite typing of Candida parapsilosis sensu stricto isolates |
| title_short | Design and validation of a multiplex PCR protocol for microsatellite typing of Candida parapsilosis sensu stricto isolates |
| title_sort | design and validation of a multiplex pcr protocol for microsatellite typing of candida parapsilosis sensu stricto isolates |
| topic | Candida parapsilosis sensu stricto Genotyping technique Microsatellites repeats Multiplex PCR Specificity Reproducibility of results |
| url | http://link.springer.com/article/10.1186/s12864-018-5065-3 |
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