Rapid Identification of Neisseria gonorrhoeae, Chlamydia trachomatis, and Ureaplasma urealyticum by RNA Amplification...

Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT), and Ureaplasma urealyticum (UU) are common pathogens associated with sexually transmitted diseases (STDs). NG is the causative agent of gonorrhea, which can result in various disorders of the genitourinary system, including epididymitis, prostatitis, cervicitis, and infertility. CT is a microorganism responsible for non-gonococcal urethritis and pelvic inflammatory disease. UU is associated with non-gonococcal urethritis, cervicitis, prostatitis, and infertility. Current molecular technologies for the analysis of NG, CT, and UU are often complex and time-consuming. Rapid molecular testing for these pathogens may facilitate the early diagnosis of STDs. The RNA amplification lateral flow assay (RGT) is a method that does not require nucleic acid extraction. In this process, samples are lysed to release nucleic acid fragments, which are then amplified through reverse transcription and transcription. The amplified RNA product is recognized and captured by specific probes, forming an RNA-detection probe-gold probe complex that can be immobilized on a nitrocellulose membrane via lateral flow to produce visible bands. The entire procedure takes less than 1 h. The aim of this study was to evaluate the efficacy of RGT for the rapid detection of NG, CT, and UU. A total of 1,416 samples were included. The consistency of NG, CT, and UU when analyzed by RGT compared to the polymerase chain reaction (PCR) fluorescence probing method was 99.03% (307/310), 99.38% (159/160), and 99.02% (303/306), respectively. The consistency of NG, CT, and UU when analyzed by RGT compared to gene sequencing was 99.17% (238/240), 98.95% (188/190), and 98.30% (173/176), respectively. Compared with the pathogen isolation culture method, the detection rates for NG, CT, and UU assays were 100.00% (9/9, 17/17, 8/8). The high sensitivity and specificity, ease of use, and reduced time requirements make this assay ideal for the rapid and accurate detection of NG, CT, and UU.