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Pooling as a strategy for the timely diagnosis of soil-transmitted helminths in stool: value and reproducibility
Wright, J Pilotte, N Chooneea, D Schär, F Truscott, JE Dunn, JC Gardiner, I Williams, SA
Wright, J
Pilotte, N
Chooneea, D
Schär, F
Truscott, JE
Dunn, JC
Gardiner, I
Williams, SA
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2019-09-16
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2019-12-03
Subject Terms
Breakpoint of transmission
Cost analysis of pooling
Pooling
Soil-transmitted helminths
Stool samples
qPCR-based diagnostics
Cost analysis of pooling
Pooling
Soil-transmitted helminths
Stool samples
qPCR-based diagnostics
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Abstract
Background
The strategy of pooling stool specimens has been extensively used in the field of parasitology in order to facilitate the screening of large numbers of samples whilst minimizing the prohibitive cost of single sample analysis. The aim of this study was to develop a standardized reproducible pooling protocol for stool samples, validated between two different laboratories, without jeopardizing the sensitivity of the quantitative polymerase chain reaction (qPCR) assays employed for the detection of soil-transmitted helminths (STHs). Two distinct experimental phases were recruited. First, the sensitivity and specificity of the established protocol was assessed by real-time PCR for each one of the STHs. Secondly, agreement and reproducibility of the protocol between the two different laboratories were tested. The need for multiple stool sampling to avoid false negative results was also assessed. Finally, a cost exercise was conducted which included labour cost in low- and high-wage settings, consumable cost, prevalence of a single STH species, and a simple distribution pattern of the positive samples in pools to estimate time and money savings suggested by the strategy.
Results
The sensitivity of the pooling method was variable among the STH species but consistent between the two laboratories. Estimates of specificity indicate a ‘pooling approach’ can yield a low frequency of ‘missed’ infections. There were no significant differences regarding the execution of the protocol and the subsequent STH detection between the two laboratories, which suggests in most cases the protocol is reproducible by adequately trained staff. Finally, given the high degree of agreement, there appears to be little or no need for multiple sampling of either individuals or pools.
Conclusions
Our results suggest that the pooling protocol developed herein is a robust and efficient strategy for the detection of STHs in ‘pools-of-five’. There is notable complexity of the pool preparation to ensure even distribution of helminth DNA throughout. Therefore, at a given setting, cost of labour among other logistical and epidemiological factors, is the more concerning and determining factor when choosing pooling strategies, rather than losing sensitivity and/or specificity of the molecular assay or the method.
Citation
Papaiakovou, M., Wright, J., Pilotte, N. et al. Pooling as a strategy for the timely diagnosis of soil-transmitted helminths in stool: value and reproducibility. Parasites Vectors 12, 443 (2019) doi:10.1186/s13071-019-3693-3
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Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
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1756-3305
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openAccess