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| Establishment and Evaluation of a Rapid Detection Method for Neisseria Gonorrhoeae Nucleic Acid |
| WANG Zhi-yu, TU Bo-wen, CHEN Hong-yan |
| School of Life Sciences Fudan University, Shanghai, 200082 |
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Abstract 【Objective】 To establish a Neisseria gonorrhoeae (NG) nucleic acid rapid test method and assess the analytic sensitivity and specificity and detection consistency with quantitative PCR method. 【Methods】 The rapid detection method of Neisseria gonorrhoeae nucleic acid was established by RPA-LFA; the sensitivity and specificity of the method were evaluated by using the national reference for Neisseria gonorrhoeae nucleic acid detection; sixty-one samples of clinical genital secretions were detected by fluorescence quantitative PCR kit and RPA-LFA respectively, and the consistency of the detection results was evaluated.【Results】 The sensitivity of RPA-LFA method was 104 cells/mL, the specificity was good, 10 different Neisseria gonorrhoeae strains could be detected, 10 non Neisseria gonorrhoeae strains could not be detected. The consistency between RPA-LFA method and fluorescent quantitative PCR method was high (Kappa=0.97), the positive detection rate of RPA-LFA method was 52.5% (32/61) and that of fluorescent quantitative PCR method was 54.1% (33/61). The sensitivity and specificity of rpa-lfa were 96.97% and 100.00% respectively, and the flow time of real-time PCR for single sample was 120±20min, significantly longer than 35±5min of RPA-LFA (P<0.05).【Conclusion】 The RPA-LFA method for rapid detection of Neisseria gonorrhoeae nucleic acid is successfully established. The method has high consistency with the fluorescent quantitative PCR method, and does not need complex and precise temperature control equipment. The interpretation of detection results is simple, which shortens the detection time of Neisseria gonorrhoeae nucleic acid and reduces the cost.
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Received: 27 June 2019
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[1] Van Dyck E, Ieven M, Pattyn S, et al. Detection of chlamydia trachomatis and neisseria gonorrhoeae by enzyme immunoassay, culture, and three nucleic acid amplification tests[J].Clin Microb,2001, 39(5): 1751-1756.
[2] 许心美, 陈培明. 无症状性病感染者及防治对策[J].中国皮肤性病学杂志, 2002, 16(6):416.
[3] Jephcott AE. Microbiological diagnosis of gonorrhoea[J].Genitourin Med,1997, 73(4): 245-252.
[4] 许菁. 实时荧光定量PCR法检测NG、CT和UU的临床意义[J].实用预防医学, 2008, 15(6): 1967-1968.
[5] Piepenburg O, Williams CH, Stemple DL, et al. DNA detection using recombination proteins[J].PLoS Biol,2006, 4(7): e204.
[6] Wu YD, Xu MJ, Wang QQ, et al. Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for detection of Toxoplasma gondii in the environment[J]. Vet Parasitol,2017, 243(3):199-203.
[7] Kr lov K, Frolova J, Tudoran O, et al. Sensitive and rapid detection of Chlamydia trachomatis by recombinase polymerase amplification directly from urine samples[J].J Mol Diagn,2014, 16(1): 127-135.
[8] Crannell ZA, Cabada MM, Castellanos-Gonzalez A, et al. Recombinase polymerase amplification-based assay to diagnose Giardia in stool samples[J].Am J Trop Med Hyg,2015, 92(3): 583-587.
[9] Yin F, Liu J, Liu A, et al. Rapid diagnosis of Theileria annulata by recombinase polymerase amplification combined with a lateral flow strip (LF-RPA) in epidemic regions[J].Vet Parasitol,2017, 237: 125-129.
[10] Yang Y, Qin X, Zhang X, et al. Development of real-time and lateral flow dipstick recombinase polymerase amplification assays for rapid detection of goatpox virus and sheeppox virus[J].Virol J,2017, 14(1): 131.
[11] Hou P, Zhao G, Wang H, et al. Development of a recombinase polymerase amplification combined with lateral-flow dipstick assay for detection of bovine ephemeral fever virus[J].Mol Cell Probes,2018, 38(1)): 31-37.
[12] Yang Y, Qin X, Wang G, et al. Development of an isothermal amplification-based assay for rapid visual detection of an Orf virus[J].Virol J,2016, 13(1): 46. |
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2020, Vol. 37 
