Year: 2026
Type: article
Abstract: Background: Disease surveillance is critical for WHO’s Global Programme to Eliminate Lymphatic Filariasis (LF), but current field-ready diagnostics exhibit insufficient sensitivity and/or specificity in low-endemic settings. We aim to develop an ultra-sensitive and field-friendly CRISPR/Cas13-based SHERLOCK (Specific High-sensitivity Enzymatic Reporter unLOCKing) tool for point-of-care detection of Wuchereria bancrofti infection. Methods: The SHERLOCK assay was performed in two-step and one-pot formats with sensitivity compared to qPCR. Assays were validated using 149 pooled mosquito and 185 human blood samples from Samoa, which were previously tested for LF antigen using WHO-recommended Filariasis Test Strips (FTS) and microscopically examined for microfilariae (Mf). We also used lyophilised one-pot SHERLOCK master mix and assessed its thermostability after storage at 25°C, 4°C and -20°C. Findings: The W. bancrofti SHERLOCK assays achieved a detection limit of 0.1 fg/μL gBlock DNA and showed no cross-reaction with five other filarial species. For both mosquito and human samples, the two-step SHERLOCK assay reached 100% sensitivity and specificity, while one-pot SHERLOCK showed 94.0-96.4% sensitivity and 100% specificity. The lyophilised one-pot SHERLOCK mix maintained a detection limit of 10 fg/μL W. bancrofti gBlock after 10 days at 25°C, indicating reduced reliance on cold-chain for reagent storage and transport. However, when testing human blood samples with lyophilised SHERLOCK, reduced sensitivity (59.7%) was observed. Interpretation: This CRISPR-based platform offers a promising field-ready next-generation approach to LF diagnostics, potentially addressing current limitations, enhancing surveillance and elimination efforts.
Source: Emerging Microbes & Infections
Institutions: QIMR Berghofer Medical Research Institute, James Cook University, Federation University, Queensland University of Technology, Australian Institute of Tropical Health and Medicine, Australian Defence Force Malaria and Infectious Disease Institute, La Trobe University, The University of Melbourne, The University of Queensland, Chinese Academy of Sciences, National University of Samoa
Language: English
Subfield: Infectious Diseases
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $2,643
OpenAlex ID: W7154910109