Field Evaluation of a Loop-Mediated Isothermal Amplification (LAMP) Platform for the Detection of Schistosoma japonicum Infection in Oncomelania hupensis Snails.

Zhi-Qiang Qin,Yin-Long Li,Shan Lv,Shi-Zhu Li,Ting Feng,Robert Bergquist,Ying-Jun Qian,Li-Juan Zhang,Chao Lv,Jing Xu,Xiao-Nong Zhou

doi:10.3390/tropicalmed3040124

Abstract

Schistosoma infection in snails can be monitored by microscopy or indirectly by sentinel mice. As both these approaches can miss infections, more sensitive tests are needed, particularly in low-level transmission settings. In this study, loop-mediated isothermal amplification (LAMP) technique, designed to detect a specific 28S ribosomal Schistosoma japonicum (Sj28S) gene with high sensitivity, was compared to microscopy using snail samples from 51 areas endemic for schistosomiasis in five Chinese provinces. In addition, the results were compared with those from polymerase chain reaction (PCR) by adding DNA sequencing as a reference. The testing of pooled snail samples with the LAMP assay showed that a dilution factor of 1/50, i.e., one infected snail plus 49 non-infected ones, would still result in a positive reaction after the recommended number of amplification cycles. Testing a total of 232 pooled samples, emanating from 4006 snail specimens, showed a rate of infection of 6.5%, while traditional microscopy found only 0.4% positive samples in the same materials. Parallel PCR analysis confirmed the diagnostic accuracy of the LAMP assay, with DNA sequencing even giving LAMP a slight lead. Microscopy and the LAMP test were carried out at local schistosomiasis-control stations, demonstrating that the potential of the latter assay to serve as a point-of-care (POC) test with results available within 60–90 min, while the more complicated PCR test had to be carried out at the National Institute of Parasitic Diseases (NIPD) in Shanghai, China. In conclusion, LAMP was found to be clearly superior to microscopy and as good as, or better than, PCR. As it can be used under field conditions and requires less time than other techniques, LAMP testing would improve and accelerate schistosomiasis control.

Highlights

Schistosomiasis, one of the neglected tropical diseases (NTDs), is a public health problem caused by one of six species of the Schistosoma parasite that affects >200 million people in Africa, South America and Southeast Asia
As part of this study, we explored the application of loop-mediated isothermal amplification (LAMP) using pooled snail samples, i.e., instead of testing each snail separately, we combined snails, never in numbers exceeding 50 snails per pooled sample, based on preliminary dilution tests
To make sure that the LAMP test had the required specificity, it was applied to 20 pools produced from a total of 1000 individual snails that had previously been investigated by microscopy

Summary

Introduction

Schistosomiasis, one of the neglected tropical diseases (NTDs), is a public health problem caused by one of six species of the Schistosoma parasite that affects >200 million people in Africa, South America and Southeast Asia. In 2004, a new integrated control strategy was introduced [5,6] involving reduction of infection sources by fencing off transmission sites, the replacement of water buffaloes (an important reservoir) with tractors for agricultural work and improved sanitation via access to clean water and latrines. These approaches have markedly reduced the infection rate in humans, domestic animals and the intermediate snail host [7]. To sustain the success achieved with regard to schistosomiasis elimination in China, highly sensitive snail-monitoring systems capable of assessing residual transmission in real time are needed

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