Comparison of culture-based, vital stain and PMA-qPCR methods for the quantitative detection of viable hookworm ova

Abstract

Accurate quantitative measurement of viable hookworm ova from environmental samples is the key to controlling hookworm re-infections in the endemic regions. In this study, the accuracy of three quantitative detection methods [culture-based, vital stain and propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR)]was evaluated byenumerating 1,000±50Ancylostoma caninum ova in the laboratory. The culture-based method was able to quantify an average of 397±59 viable hookwormova. Similarly, vital stain and PMA-qPCRmethods quantified 644± 87 and 587±91 viable ova, respectively. The numbers of viable ova estimated by the culture-based method were significantly (P< 0.05) lower than vital stain and PMA-qPCR methods. Therefore, both PMA-qPCR and vital stain methods appear to be suitable for the quantitative detection of viable hookwormova. However, PMA-qPCRwould be preferable over the vital stain method in scenarios where ova speciation is needed.