Comparison of deferred and bedside culture of Neisseria gonorrhoeae: a study to improve the isolation of gonococci for antimicrobial susceptibility testing
Background and Objectives: Antimicrobial resistance of Neisseria gonorrhoeae is globally spread and threatening. Culturing of N. gonorrhoeae is the only method to collect live isolates for investigation antimicrobial resistance profile. Therefore, quality assessment of N. gonorrhoeae culture is essential for successful isolation of gonococci. This study was conducted to evaluate deferred and bedside culture of N. gonorrhoeae depending on the year season and temperature condition of transport media temporary storage.
Materials and Methods: Urogenital swabs from 46 symptomatic heterosexual patients with gonorrhoea and subculture of N. gonorrhoeae in 46 suspensions in concentrations 1.5 × 108 CFU/ml were subjected to the study. Non-nutritive transporting medium Amies Agar Gel Medium with charcoal (Copan Diagnostics Inc., Brescia, Italy) was used for deferred culture and selective Chocolate agar TM+PolyViteX VCAT3 (BioMérieux, Marcy-l'Étoile, France) for both tested methods of culture.
Results: The specificity of both bedside and deferred methods of culture was 100%. The sensitivity of deferred culture was higher than of bedside culture (82.6% vs 47.8%, p<0.0005). Deferred culture showed significantly higher sensitivity comparing to bedside culture in summer (100% vs 50%, p=0.003), and comparably the same as for bedside culture in autumn, winter and spring.
Conclusion: The viability of N. gonorrhoeae subcultures was significantly higher in refrigerated samples from transport media than from ambient one after exposition from 48 to 96 hours. Optimal viability of N. gonorrhoeae was observed when transport swabs were kept refrigerated up to 48 h (73.9-93.5%) or ambiently – up to 24 h (87%). Updating laboratory guidelines regarding sampling and timely specimen processing might improve gonococcal culture performance.
2. Unemo M, Seifert HS, Hook EW 3rd, Hawkes S, Ndowa F, Dillon JR. Gonorrhoea. Nat Rev Dis Primers 2019; 5:79.
3. European Centre for Disease Prevention and Control. Disease data from ECDC Surveillance Atlas – gonorrhoea. https://www.ecdc.europa.eu/en/gonorrhoea/surveillance-and-disease-data/disease-data-atlas
4. Tsevat DG, Wiesenfeld HC, Parks C, Peipert JF. Sexually transmitted diseases and infertility. Am J Obstet Gynecol 2017; 216:1-9.
5. Koval HD, Chopyak VV, Kamyshnyi OM, Kurpisz MK. Transcription regulatory factor expression in T-helper cell differentiation pathway in eutopic endometrial tissue samples of women with endometriosis associated with infertility. Cent Eur J Immunol 2018; 43:90-96.
6. Cole MJ, Quinten C, Jacobsson S, Michaela Day, Andrew J, Amato-Gauci, et al. The European gonococcal antimicrobial surveillance programme (Euro-GASP) appropriately reflects the antimicrobial resistance situation for Neisseria gonorrhoeae in the European Union/European Economic Area. BMC Infect Dis 2019;19:1040.
7. Unemo M, Clarke E, Boiko I, Patel C, Patel R. Adherence to the 2012 European gonorrhoea guideline in the WHO European region according to the 2018-19 international union against sexually transmitted infections European collaborative clinical group gonorrhoea survey. Int J STD AIDS 2019; 31:095646241987927.
8. Sánchez-Busó L, Golparian D, Corander J, Grad YH, Ohnishi M, Flemming R, et al. The impact of antimicrobials on gonococcal evolution. Nat Microbiol 2019; 4:1941-1950.
9. Tapsall JW. Antibiotic resistance in Neisseria gonorrhoeae. Clin Infect Dis 2005;41 Suppl 4:S263-S268.
10. Unemo M, Ballard R, Ison C, Lewis D, Ndowa F, Peeling R. Laboratory diagnosis of sexually transmitted infections, including human immunodeficiency virus. Geneva: World Health Organization Document Production Services, 2013. Available at: https://apps.who.int/iris/bitstream/handle/10665/85343/9789241505840_eng.pdf?sequence=1&isAllowed=y
11. Domeika M, Frankenberg A, Svetashov O, Kovalenko J, Bojko I, Glushok O, et al. P2.022 Laboratory diagnosis of genital Neisseria gonorrhoeae infections in three regions of Ukraine. Sex Transm Infect 2013; 89(Suppl 1):A94-A94.
12. Serra-Pladevall J, Caballero E, Roig G, Juvé R, Barbera MJ, Andreu A. Comparison between conventional culture and NAATs for the microbiological diagnosis in gonococcal infection. Diagn Microbiol Infect Dis 2015; 83:341-343.
13. Barlow D. Culture of the gonococcus: a reliable gold standard? Sex Transm Infect 2017; 93:178.
14. Glebova EE, Horachuk VV, Verba EA. Organization of dermatovenerologic medical care: domestic realities and world practice (review of literature). Wiad Lek 2019;72:2187-2191.
15. Boiko I, Akimova V, Mazur L, Savchenko I, Kohut I, Krynytska I. The clinico-epidemiological profile of patients with gonorrhoea and challenges in the management of Neisseria gonorrhoeae infection in an STI clinic, Ternopil, Ukraine (2013-2018). J Med Life 2020;13:75-81.
16. Jacobsson S, Boiko I, Golparian D, Blondeel K, Kiarie J, Toskin I, et al. WHO laboratory validation of Xpert® CT/NG and Xpert® TV on the GeneXpert system verifies high performances. APMIS 2018;126:907-912.
17. Boiko I, Golparian D, Krynytska I, Unemo M. High prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae and particularly Trichomonas vaginalis diagnosed using US FDA-approved Aptima molecular tests and evaluation of conventional routine diagnostic tests in Ternopil, Ukraine. APMIS 2019; 127:627-634.
18. Domeika M, Frankenberg A, Svetashov O, Kovalenko J, Bojko I, Glushok O, et al. P2.022 Laboratory diagnosis of genital Neisseria gonorrhoeae infections in three regions of Ukraine. Sex Transm Infect 2013; 89(Suppl 1):A94-A94.
19. Unemo M, Savicheva A, Budilovskaya O, Sokolovsky E, Larsson M, Domeika M. Laboratory diagnosis of Neisseria gonorrhoeae in St Petersburg, Russia: inventory, performance characteristics and recommended optimisations. Sex Transm Infect 2006; 82:41-44.
20. Cole MJ, Quaye N, Jacobsson S, Day M, Fagan E, Ison C, et al. Ten years of external quality assessment (EQA) of Neisseria gonorrhoeae antimicrobial susceptibility testing in Europe elucidate high reliability of data. BMC Infect Dis 2019; 19:281.
21. Olsen CC, Schwebke JR, Benjamin WH Jr, Beverly A, Waites KB. Comparison of direct inoculation and Copan transport systems for isolation of Neisseria gonorrhoeae from endocervical specimens. J Clin Microbiol 1999; 37:3583-3585.
22. Radebe F, Gumede L, Ricketts C, Vezi A, Maseko V, Lewis DA. The evaluation and comparison of two transport media for the growth, holding and transport of Neisseria gonorrhoeae. Int J Med Biomed Sci 2014; 2:39-45.
23. Serra-Pladevall J, Gulin Blanco C, Vila Olmo N, Arjona Camacho P, Andreu Domingo A. Preservation of Neisseria gonorrhoeae: should swabs be refrigerated or not?: Neisseria gonorrhoeae preservation. J Microbiol Methods 2017;145:37-39.
24. Farhat SE, Thibault M, Devlin R. Efficacy of a swab transport system in maintaining viability of Neisseria gonorrhoeae and Streptococcus pneumoniae. J Clin Microbiol 2001; 39:2958-2960.
25. Boiko I, Golparian D, Krynytska I, Bezkorovaina H, Frankenberg A, Onuchyna M, et al. Antimicrobial susceptibility of Neisseria gonorrhoeae isolates and treatment of gonorrhoea patients in Ternopil and Dnipropetrovsk regions of Ukraine, 2013-2018. APMIS 2019; 127:503-509.