Validation of a quantitative Eimeria spp. PCR for fresh droppings of broiler chickens.

Validation of a quantitative Eimeria spp. PCR for fresh droppings of broiler chickens.
Avian Pathol. 2017 Jun 05;:1-25
Authors: Peek HW, Ter Veen C, Dijkman R, Landman WJM
Abstract
A quantitative Polymerase Chain Reaction (qPCR) for the seven chicken Eimeria spp. was modified and validated for direct use on fresh droppings. The analytical specificity of the qPCR on droppings was 100%. Its analytical sensitivity (non-sporulated oocysts/g droppings) was 41 for E. acervulina, ≤2,900 for E. brunetti, 710 for E. praecox, 1,500 for E. necatrix, 190 for E. tenella, 640 for E. maxima and 1,100 for E. mitis. Field validation of the qPCR was done using droppings with non-sporulated oocysts from 19 broilers flocks. To reduce the number of qPCR tests five grams of each pooled sample per time point were blended into one 'mixed' sample. Comparison of the OPG-counting method with the qPCR using pooled samples (n = 1180) yielded a Pearson's correlation coefficient of 0.78 (95% CI: 0.76 - 0.80) and a Pearson's correlation coefficient of 0.76 (95% CI: 0.70 - 0.81) using mixed samples (n = 236). Comparison of the average of the OPG-counts of the five pooled samples with the mixed sample per time point (n = 236) showed a Pearson's correlation coefficient (R) of 0.94 (95% CI: 0.92 - 0.95) for the OPG-counting method and 0.87 (95% CI: 0.84 - 0.90) for the qPCR. This indicates that mixed samples are practically equivalent to the mean of five pooled sampl...

Source: Avian Pathology - June 5, 2017 Category: Pathology Authors: Peek HW, Ter Veen C, Dijkman R, Landman WJM Tags: Avian Pathol Source Type: research