Estimation of the relative sensitivity of qPCR analysis using pooled samples

Ana Muniesa; Chelo Ferreira; Héctor Fuertes; Nabil Halaihel; Ignacio de Blas

doi:10.1371/journal.pone.0093491

Estimation of the relative sensitivity of qPCR analysis using pooled samples

PLoS One. 2014 Apr 10;9(4):e93491. doi: 10.1371/journal.pone.0093491. eCollection 2014.

Authors

Ana Muniesa¹, Chelo Ferreira², Héctor Fuertes¹, Nabil Halaihel¹, Ignacio de Blas¹

Affiliations

¹ Department of Animal Pathology, Facultad de Veterinaria, Universidad de Zaragoza, Zaragoza, Spain.
² Department of Applied Mathematics, Facultad de Veterinaria, Universidad de Zaragoza, Zaragoza, Spain.

Abstract

The high sensitivity of qPCR makes it a desirable diagnostic method in epidemiological surveillance programs. However, due to high costs, the use of pooling has been suggested. In this paper, an algorithm based on the Montecarlo method has been designed and implemented. The algorithm had been tested in many different situations, and finally it was validated with a real dataset. Moreover, based on the results obtained and depending on pooling conditions, a drastic decrease of sensitivity is observed.

MeSH terms

Algorithms
Animals
Bacterial Infections / diagnosis*
Humans
Models, Statistical
Monte Carlo Method
Prevalence
Probability
Real-Time Polymerase Chain Reaction / methods*
Regression Analysis
Reproducibility of Results
Sensitivity and Specificity

Grants and funding

Ministerio de Educación of Spain is funding a FPU/MEC doctorate fellowship for Ana Muniesa. Also the Departamento de Industria e Innovación del Gobierno de Aragón, Fondo Social Europeo and Dirección General de Ciencia y Tecnología (REF. MTM2010-21037) provides financial support to Research Groups A35 and T53 for their basic running costs. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.