Abstract

Herein, a new molecular diagnosis assay for rapid detection of bacterial DNA was established, which combined single primer isothermal amplification (SPIA) with rapid isothermal detection assay (RIDA). Using this method, we could rapidly identify Group BStreptococcus DNA with high sensitivity and specificity, the detection limit of the new method was 10²copy/mL. The whole assay was processed in one tube and the result was determined by naked eyes under ultraviolet radiation without cap-opening, meanwhile, as a linear amplification system, the products cannot be used as the amplification target, thus, the aerosol contamination was further decreased. This novel method shows excellent sensitivity, specificity and conveniences, which is easily operated in the elementary medical organizations and resource-limited areas.

Abbreviation

GBS, Group B Streptococcus; SPIA, Single primer isothermal amplification; PCR, polymerase chain reaction; RIDA, rapid isothermal detection assay;RP, reporting probe; Tm,melting temperature; NASBA,nucleic acid sequence-based amplification.