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

Table 1 Main characteristics, pros and cons between four C. parapsilosis sensu stricto microsatellite genotyping protocols published in the literature compared and the one described in this work (N = 5)

Characteristic	Sabino et al. [5]	Diab-Elschahawi et al. [6]	Reiss et al. [7]	Vaz et al. [8]	Trobajo-Sanmartín et al. (This study)
PCR scheme used	Singleplex^b	Multiplex	Multiplex	Multiplex	Multiplex
Redesign of the original primers	NA	Yes	Yes	Yes	No
Number of primers pairs used for each PCR reaction	1	3	1	4	4
Total number of PCR reactions needed for a complete STR analysis	3	2	5	1	1
Number of dyes used in each genotyping reaction	3	3	1	3	4
Evaluation of the sensitivity and specificity^a	NA	No	No	No	Yes
Evaluation of the microsatellite informativeness parameters	Yes	Unknown^c	Unknown^c	Unknown^b	Yes
Total time elapsed to obtained results (in minutes)	138	79	197	138	79
Approximate cost^d estimated of the primer pairs used in each protocol (in euros)	5,60	15,72	7,00	9,26	12,76
Approximate total cost^d estimated for the complete STR analysis of one sample (in euros)	15,58	22,53	20,77	14,00	17,45

NA Not applicable
^aTaking the original protocol published by Sabino and coworkers as the gold standard
^bOne of the four reactions could be multiplexed using three different dyes
^cProbably the authors did the evaluation of the microsatellite informativeness parameters, but there is no clear reference to that subject in their manuscript
^dPrices are referred to Spain. We included the DNA extraction, AmpliTaq Gold® PCR kit with magnesium chloride and PCR buffer, PCR plate (96-well) and primer prices to estimate the total cost per sample analyzed using each protocol

ISSN: 1471-2164