Progress in loop-mediated isothermal amplification assay for detection of Schistosoma mansoni DNA: towards a ready-to-use test

Abstract

Schistosomiasis is one of the most prevalent Neglected Tropical Disease, affecting approximately 250 million people worldwide. Schistosoma mansoni is the most important species causing human intestinal schistosomiasis. Despite significant efforts in recent decades, the global disease burden of schistosomiasis remains extremely high. This could partly be attributed to the absence of accurate diagnostic tools, primarily in endemic areas. Loop-mediated isothermal amplification (LAMP) is increasingly used in molecular diagnostics as a field-friendly alternative to many other complex molecular methods and it has been proposed as an ideal candidate for revolutionizing point-of-care molecular diagnostics. In a previous work, a LAMP-based method to detect S. mansoni DNA (SmMIT-LAMP) was developed by our research group for early diagnosis of active schistosomiasis in an experimental infection murine model. The SmMIT-LAMP has been further successfully evaluated in both human stool and snail samples and, recently, in human urine samples. In this study, we developed an important improvement for SmMIT-LAMP molecular assay, transforming it into a cold maintenance dry format suitable for potentially manufacturing as kit for ready-to-use for schistosomiasis diagnosis. This procedure could be applied to create dry LAMP kits for a laboratory setting and for diagnostic applications for other neglected tropical diseases.

Molecular Markers for Detecting Schistosoma Species by Loop-Mediated Isothermal Amplification

“…The infection is endemic in 78 countries, mainly in tropical and subtropical areas, although it predominates in Sub-Saharan Africa where more than 80% of the cases occur, leading to around 280,000 deaths annually. The Global Burden of Disease study attributed 1.43 million disability-adjusted life years (DALYs) to it in 2017 [ 2 – 5 ].…”

Loop-Mediated Isothermal Amplification as Point-of-Care Diagnosis for Neglected Parasitic Infections

“…The authors modified the protocol for obtaining this limit, increasing incubation time from 60 to 120 minutes; this improved analytical sensitivity from 1 fg to 0.01 fg and increasing incubation time did not affect test specificity. Using this set of primers which had already been tested in urine [152], faeces [158] and snails [155], the research group improved the protocol, transforming it into a cold maintenance dry format, suitable for potentially manufacturing as a kit for ready-to-use for schistosomiasis diagnosis [150]. Price et al [151] amplified the Sm1-7 sequence for evaluating the effect of extraction on LAMP performance, finding greater sensitivity and specificity when samples were extracted with a LAMP-Qiagen kit (100% sensitivity: 96-100% 95% CI; 100% specificity: 59-100% 95% CI), compared to using PCR-Qiagen, PCR_LAMP-PURE, LAMP_LAMP_PURE kits.…”

Core-Shell Beads as Microreactors for Phylogrouping of E. coli Strains

“…For example, agarose gel-based DNA separation technique is a necessary step for both assays. Conventional agarose gel-based technique has limitations such as inter-laboratory difference in explaining banding patterns, time-consuming and complex processes, and the need for skilled personnel as well as expensive equipment [25,26]. Although some previously reported systems such as labelled probes have been also used for direct target gene identification without the need for electrophoresis, the implementation of these probe-based sensors is time consuming and requires multiple PCR product handling steps [27].…”