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Journal of Clinical Microbiology, August 2007, p. 2743-2744, Vol. 45, No. 8
0095-1137/07/$08.00+0     doi:10.1128/JCM.00344-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

In-House Growth-Promoting Transport System for Neisseria gonorrhoeae{triangledown}

Dharmendra Sharma,1 Sunil Sethi,1* Swami Das Mehta,2 and Meera Sharma1

Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research,1 Department of Dermatology and Venerology, STD Polyclinic Sector 22, Chandigarh, India2

Received 13 February 2007/ Returned for modification 22 March 2007/ Accepted 16 May 2007


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ABSTRACT
 
Eno powder (GlaxoSmithKline), an antacid preparation readily available over the counter, was used instead of a CO2 generator for the growth of 15 strains of Neisseria gonorrhoeae obtained from men with urethritis. Due to its easy accessibility and low cost, Eno powder can be useful in developing countries for transporting clinical specimens from resource-poor peripheral labs to reference laboratories.


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TEXT
 
Despite a sharp decline in the incidence of gonorrhea in developed countries during the last decade, gonococcal infection remains a significant public health problem in developing countries (9, 10). Timely and accurate laboratory detection of gonorrhea is of major public health importance because untreated gonococcal infection may lead to several complications, especially in women, e.g., ectopic pregnancy, pelvic inflammatory disease, and tubal infertility (4, 5). Moreover, gonorrhea increases the risk of human immunodeficiency virus infection. Neisseria gonorrhoeae is a nutritionally fastidious organism that requires an enriched medium in a moist environment of 35 to 37°C with an atmosphere of 3 to 5% CO2 for growth. It does not grow well in the presence of commensal organisms (6, 7, 8).

Ideally, clinical specimens should be directly inoculated onto a suitable gonococcal-culture selective medium and immediately incubated at 36°C in 5 to 10% CO2. However, the necessary facilities are not available in most venereal disease clinics. To circumvent this need, specialized transport systems like the Gono-Pak (BBL), the JEMBEC (BBL), and the Bio-Bag Type C (Becton Dickinson) systems have been developed (1). These systems work by producing a CO2-rich environment. In the Gono-Pak system, a modified Thayer-Martin medium plate is placed in a ziplock plastic bag along with a CO2-generating tablet. The JEMBEC system is made up of a rectangular plastic tray which contains modified Thayer-Martin medium (1). The tray has a well into which a CO2-generating tablet is placed. The tray is placed inside a ziplock plastic bag (1). The disadvantage of the Gono-Pak and JEMBEC systems is that the CO2 generation by the tablet is dependent on the release of moisture from the medium. Thus, the efficacy of these systems is dependent on the freshness of the medium. The Bio-Bag Type C system eliminates dependence on the freshness of the medium. In this system, CO2 is produced by a CO2 generator which contains a 130-mg sodium bicarbonate tablet and a 0.5-ml ampoule of 1.56 N hydrochloric acid, packaged in a ziplock plastic bag (1). The ampoule is crushed to generate CO2. The cost of each of these systems would make them prohibitive in small labs of developing countries like India. Here, we are sharing our experience with an in-house transport system which can be made readily in any microbiology laboratory of a developing country.

Our system consists of a ziplock plastic bag in which a plate of modified New York City medium is placed. For the generation of CO2 in the bag, Eno powder (GlaxoSmithKline, Asia Pvt. Ltd.), an antacid preparation readily available over the counter, is used. Eno powder, which is available in air-sealed 5-gram sachets, contains sodium bicarbonate and citric acid in solid form, which generate CO2 after becoming wet (3). Our system also contains a wet filter paper which provides moisture to the Eno powder in a gradual manner, resulting in slow and consistent generation of CO2. Additionally, the wet filter paper creates a humid environment, which supports the growth of gonococci.

The study group consisted of men who presented with symptoms of acute urethritis at the sexually transmitted disease clinics of Nehru Hospital attached to the Postgraduate Institute of Medical Education and Research, Chandigarh, India, and the STD Polyclinic Sector 22 from March 2006 to September 2006. Patients were enrolled and examined after giving oral consent. Two urethral swab specimens were obtained by gently rotating a cotton swab in the anterior urethra. The swabs were used to inoculate the modified New York City agar plates in duplicate. One plate was placed in the CO2 incubator at 37°C for up to 72 h. Another plate was placed into a ziplock plastic bag. Before the culture plate was placed in the plastic bag, a small packet of 3 g of Eno powder in a paper towel was created and affixed to the outer side of the lid of the culture plate using cellophane tape. A wet rectangular piece of filter paper was then placed inside the plastic bag. The culture plate was placed over it so that the Eno powder packet was in direct contact with the wet filter paper. The ziplock plastic bag was then sealed and incubated at 37°C in a biological oxygen demand incubator.

The plates were examined daily for up to 72 h for colonies typical of N. gonorrhoeae. Isolates which were oxidase-producing gram-negative diplococci were confirmed to be N. gonorrhoeae by the rapid carbohydrate utilization test (2). Specimens from a total of 20 patients were tested in this manner, and 15 of these specimens were culture positive for N. gonorrhoeae. There was complete agreement between the results with the plates incubated in a CO2 incubator and those incubated in the Eno system. Our study shows that the Eno system works as well as the CO2 incubator to support the growth of gonococci. Due to the easy accessibility and the low cost ($0.04 per sachet) of Eno powder, it may be useful for transporting gonococcal specimens from resource-poor peripheral labs to reference laboratories. The Eno system also has the advantage of a consistent release of CO2 and is not dependent upon the freshness (moisture content) of the medium, as are the Gono-Pak and JEMBEC systems.


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FOOTNOTES
 
* Corresponding author. Mailing address: Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research, Chandigarh, India 160012. Phone: 91 172 2755161. Fax: 91 172 2744401. E-mail: sunilsethi10{at}hotmail.com Back

{triangledown} Published ahead of print on 30 May 2007. Back


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REFERENCES
 
    1
  1. Carlson, B. L., M. S. Haley, N. A. Tisei, and W. M. McCormack. 1980. Evaluation of four methods for isolation of Neisseria gonorrhoeae. J. Clin. Microbiol. 12:301-303.[Abstract/Free Full Text]
    2. 2
  2. Fallon, R. J., and H. Young. 1996. Neisseria, Moraxella, Acinetobacter, p. 263-297. In J. G. Collee, B. P. Marmion, A. G. Fraser, and A. Simmons (ed.), Mackie and McCartney practical medical microbiology, 14th ed. Churchill Livingstone, New York, NY.
    3. 3
  3. GlaxoSmithKline. 2005. Safety data sheet number 2492. Approved/revised 09 March 2005. Brentford, Middlesex, United Kingdom.
    4. 4
  4. Hook, E. W., and K. K. Holmes. 1985. Gonococcal infections. Ann. Intern. Med. 102:229-243.[Abstract/Free Full Text]
    5. 5
  5. Jephcott, A. E. 1998. Gonorrhoea, chancroid and granuloma venereum, p. 623-640. In W. J. Hausler, Jr., and M. Sussman (ed.), Topley and Wilson's microbiology and microbial infections, 9th ed., vol. 3. Arnold, London, United Kingdom.
    6. 6
  6. Kraus, S. J., R. C. Geller, G. H. Perkins, and D. L. Rhoden. 1976. Interference of Neisseria gonorrhoeae growth by other bacterial species. J. Clin. Microbiol. 4:288-295.[Abstract/Free Full Text]
    7. 7
  7. Perry, J. L. 1997. Assessment of swab transport systems for aerobic and anaerobic organism recovery. J. Clin. Microbiol. 35:1269-1271.[Abstract]
    8. 8
  8. Sng, E. H., V. S. Rajan, K. L. Yeo, and A. J. Goh. 1982. The recovery of Neisseria gonorrhoeae from clinical specimens: effects of different temperatures, transport times, and media. Sex. Transm. Dis. 9:74-78.[Medline]
    9. 9
  9. Tapsall, J. W. 2000. Surveillance of antibiotic susceptibility of Neisseria gonorrhoeae in the WHO Western Pacific Region, 1998. WHO Western Pacific Region Gonococcal Antimocrobial Surveillance Programme. Commun. Dis. Intell. 24:1-4.[Medline]
    10. 10
  10. WHO Western Pacific Region Gonococcal Antimicrobial Surveillance Programme. 1997. Surveillance of antibiotic susceptibility of Neisseria gonorrhoeae in the WHO Western Pacific Region 1992-4. Genitourin. Med. 73:355-361.[Medline]

Journal of Clinical Microbiology, August 2007, p. 2743-2744, Vol. 45, No. 8
0095-1137/07/$08.00+0     doi:10.1128/JCM.00344-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.




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