Diagnostic microbiology and infectious disease · 2025
Evaluation of rapid point-of-care test based on qPCR for detecting Neisseria gonorrhoeae and β-lactamase to guide the use of antibiotics.
СтатьяОценочное исследованиеНазвание и аннотация: Перевод в очереди
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Zhu Shue, Huang Xuanmei, Lai Wenqing, He Yue, Liu Yixuan, Chen Rui, Liu Helu
Аннотация
OBJECTIVES: To establish a point-of-care testing (POCT), which can rapidly detect Neisseria gonorrhoeae and assess its sensitivity to β-lactam antibiotics, so as to improve the diagnosis and treatment of gonococcal infection.
METHODS: Specific primers and probes targeting the LST gene of Neisseria gonorrhoeae, the blaTEM gene and the human ACTB gene were designed for the platform . To evaluate specificity and sensitivity, nucleic acid references from Neisseria gonorrhoeae and several other pathogens were assessed. Clinical performance was assessed using secretory specimens. In addition, Neisseria gonorrhoeae isolates were evaluated using nitrocefin tests and blaTEM gene detection.
RESULTS: The platform enables the detection of Neisseria gonorrhoeae and β-lactamase within 30 minutes. The limit of detection was 1000 copies/mL, and the linear amplification range was 1.0 × 103-108 copies/mL. A total of 922 clinical samples and 63 Neisseria gonorrhoeae isolates were collected to evaluate the POCT assay. Compared with the cultured method, the positive coincidence rate of LST was 100 %, the negative coincidence rate was 98.59 %, and the total coincidence rate was 98.70 %. The total prevalence of PPNG isolates was 68.25 % and 86.05 % of these PPNG isolates possessed blaTEM. Compared with the nitrocefin tests, blaTEM detection showed a positive coincidence rate of 86.05 %, a negative coincidence rate of 75.00 %, and an overall coincidence rate of 82.54 %.
CONCLUSIONS: This study establishes a rapid and accurate molecular diagnostic method for detecting Neisseria gonorrhoeae and β-lactamase within 30 min. The POCT platform enhances diagnostic efficiency and facilitates timely and appropriate treatment of gonococcal infection.
METHODS: Specific primers and probes targeting the LST gene of Neisseria gonorrhoeae, the blaTEM gene and the human ACTB gene were designed for the platform . To evaluate specificity and sensitivity, nucleic acid references from Neisseria gonorrhoeae and several other pathogens were assessed. Clinical performance was assessed using secretory specimens. In addition, Neisseria gonorrhoeae isolates were evaluated using nitrocefin tests and blaTEM gene detection.
RESULTS: The platform enables the detection of Neisseria gonorrhoeae and β-lactamase within 30 minutes. The limit of detection was 1000 copies/mL, and the linear amplification range was 1.0 × 103-108 copies/mL. A total of 922 clinical samples and 63 Neisseria gonorrhoeae isolates were collected to evaluate the POCT assay. Compared with the cultured method, the positive coincidence rate of LST was 100 %, the negative coincidence rate was 98.59 %, and the total coincidence rate was 98.70 %. The total prevalence of PPNG isolates was 68.25 % and 86.05 % of these PPNG isolates possessed blaTEM. Compared with the nitrocefin tests, blaTEM detection showed a positive coincidence rate of 86.05 %, a negative coincidence rate of 75.00 %, and an overall coincidence rate of 82.54 %.
CONCLUSIONS: This study establishes a rapid and accurate molecular diagnostic method for detecting Neisseria gonorrhoeae and β-lactamase within 30 min. The POCT platform enhances diagnostic efficiency and facilitates timely and appropriate treatment of gonococcal infection.