Rapid detection of Human T-cell leukemia virus-1 in blood donors by a real time loop-mediated isothermal amplification assay

Abstract

Abstract It is essential to detect Human T-cell Leukemia Virus-1 (HTLV-1) accurately and quickly in order to develop prevention strategies to reduce its transmission rates. However, the development of HTLV-1 testing in blood centers (stations) laboratories faces two challenges, including expensive reagents and labor-intensive processes .Thus, for the first time, a rapid real time loop-mediated isothermal amplification (RT-LAMP) was operated to detect HTLV-1 in blood donors. Methods: As a first step, we combined the hybridization of fluorescent dye with the powerful amplification of LAMP to detect the presence of HTLV-1 plasmids containing PX gene. After that, we validated the method by performing gene test verification on blood samples and attempted to achieve one-step DNA extraction. The method was finally applied to the testing of blood donors for the presence of the HTLV-1 virus. Results: It was successful to construct an RT-LAMP to detect HTLV-1, and the level of sensitivity was as low as 34 copies of the DNA from an HTLV-1-infected blood sample, which is sufficient for a primary blood screening. It was possible to achieve high specificity in detecting HTLV-1 virus in blood donor. With simply processed blood samples, it has shown good performance in helping establish negative donor pools in areas with high rates of HTLV-1. Conclusion: With the above method, crudely treated blood samples can be detected in one step, are fluorescent, specific and sensitive, showing great potential for use in hematology and POCT.