Towards soil-transmitted helminths transmission interruption: The impact of diagnostic tools on infection prediction in a low intensity setting in Southern Mozambique

World Health Organization goals against soil-transmitted helminthiases (STH) are pointing towards seeking their elimination as a public health problem: reducing to less than 2% the proportion of moderate and heavy infections. Some regions are reaching WHO goals, but transmission could rebound if strategies are discontinued without an epidemiological evaluation. For that, sensitive diagnostic methods to detect low intensity infections and localization of ongoing transmission are crucial. In this work, we estimated and compared the STH infection as obtained by different diagnostic methods in a low intensity setting. We conducted a cross-sectional study enrolling 792 participants from a district in Mozambique. Two stool samples from two consecutive days were collected from each participant. Samples were analysed by Telemann, Kato-Katz and qPCR for STH detection. We evaluated diagnostic sensitivity using a composite reference standard. By geostatistical methods, we estimated neighbourhood prevalence of at least one STH infection for each diagnostic method. We used environmental, demographical and socioeconomical indicators to account for any existing spatial heterogeneity in infection. qPCR was the most sensitive technique compared to composite reference standard: 92% (CI: 83%– 97%) for A. lumbricoides, 95% (CI: 88%– 98%) for T. trichiura and 95% (CI: 91%– 97%) for hookworm. qPCR also estimated the highest neighbourhood prevalences for at least one STH infection in a low intensity setting. While 10% of the neighbourhoods showed a prevalence above 20% when estimating with single Kato-Katz from one stool and Telemann from one stool, 86% of the neighbourhoods had a prevalence above 20% when estimating with qPCR. In low intensity settings, STH estimated prevalence of infection may be underestimated if based on Kato-Katz. qPCR diagnosis outperformed the microscopy methods. Thus, implementation of qPCR based predictive maps at STH control and elimination programmes would disclose hidden transmission and facilitate targeted interventions for transmission interruption. Author summary Soil-transmitted helminths (STH) are parasitic worms present in over 1 billion people. Humans release STH eggs to the environment through faeces, and they become infected by egg ingestion or larvae skin penetration. The higher the number of eggs in an infection (high intensity infection) the higher the morbidity severity. For that, the World Health Organization (WHO) aims to eliminate STH as a health problem by reducing moderate and high intensity infections below 2% in 96% of endemic countries by 2030. STH infections main control strategies are mass drug administration to people at risk, and water, sanitation, and hygiene improvements. Nowadays, some regions are reaching WHO goals. Thus, tools to confirm if control strategies could be halted are needed. In this study, we selected a low intensity district in Southern Mozambique where we evaluated how different diagnostic techniques detect STH when intensity of infection is low. In addition, we also created district maps for STH estimated infection prevalence based on the different diagnostics assessed in order to identify the location of ongoing transmission. qPCR showed to be the most sensitive technique. Hence, maps based on the widely used Kato-Katz could underestimate the STH prevalence and lose hidden transmission