Abstract
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.
Highlights
Mycobacterium tuberculosis is an age-old bacterium that is difficult to eliminate
We invented a nucleic acid extraction device to overcome limitations of the regular nucleic acid kits, employed a vitrification method to overcome the ambient temperature maintenance problem for heat-unstable components, used an all-in-one package containing all the agents to solve the problems associated with sample introduction and PCR product contamination, and employed isothermal polymerase spiral reaction (PSR) method, invented by us, for nucleic acid testing, thereby circumventing the requirement of precise and expensive instruments.With all these innovations, we successfully established a low-cost, rapid, portable nucleic acid test platform, which does not require low temperature storage, professional operation skills, or precise and expensive instruments, making this method suitable for rapid nucleic acid testing in field settings or by staff of grass-root units and for household use
Tuberculosis, Bst DNA polymerase, and dNTP onto the reaction tube cover and sealed the ionic liquid at the tube bottom by using paraffin wax; based on such a packaging technology, we successfully developed an all-in-one kit to be used for rapid detection of M. tuberculosis
Summary
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. We invented a nucleic acid extraction device to overcome limitations of the regular nucleic acid kits, employed a vitrification method to overcome the ambient temperature maintenance problem for heat-unstable components, used an all-in-one package containing all the agents to solve the problems associated with sample introduction and PCR product contamination, and employed isothermal polymerase spiral reaction (PSR) method, invented by us, for nucleic acid testing, thereby circumventing the requirement of precise and expensive instruments.With all these innovations, we successfully established a low-cost, rapid, portable nucleic acid test platform, which does not require low temperature storage, professional operation skills, or precise and expensive instruments, making this method suitable for rapid nucleic acid testing in field settings or by staff of grass-root units and for household use
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
