Development and application of a rapid Mycobacterium tuberculosis detection technique using polymerase spiral reaction

Mycobacterium tuberculosis is an age-old bacterium that is difficult to eliminate. A simple and rapid diagnostic method is of great importance to prevent the spread of M . tuberculosis . Here, we developed a low-cost rapid M . tuberculosis nucleic acid detection technique, named GenePop, which enabled the storage and transport of M . tuberculosis diagnostic reagent at ambient temperatures, without the need for professional operations or expensive instrumentation. Using a vitrification method, we vitrified heat-unstable components onto the cap of a reaction tube, and placed heat-stable components at the bottom of the reaction tube by sealing them with paraffin wax. The all-in-one detection tube, when used together with our other invention—a multi-functional sample treatment tube pre-filled with a nucleic acid-releasing agent—only required three simple steps to yield results. A comparative analysis with a commercial qPCR kit for M . tuberculosis indicated that our new technique had a concordance rate of 91.6%, showing no cross-reactivity with 11 other bacteria. The complete operation time was only 65 min. It is suitable for use in field settings or by personnel in grass-root units, and is applicable in household activities, hence can be used in developing countries.