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Evaluation of Sigma Transwab® in Liquid Amies
Transport Medium for Neisseria gonorrhoeae Culture.
E. Graveling1, H. Venkatesh2, N. Banerjee3
Department of Microbiology and Infection Control,
North Laboratory, Victoria Hospital, Hayfield Road, Kirkcaldy, KY2 5AG. eleanor.graveling@nhs.net
On receipt in the laboratory the Sigma Transwab® were inoculated on to individual LCAT plates and stroked out for single colonies. The direct culture plates and the Sigma Transwab® cultured plates were then incubated in parallel Direct inoculation of LCAT (Lincomycin, Colistin sulphate, Amphotericin B, at 35 +/-2 ºC in an atmosphere of 5% CO2 for up to 48 hours. The use of Sigma Transwab® in liquid Amies transport medium obtained in Trimethoprim) agar plates has been considered technically superior for parallel with the current LCAT culture plates has provided data which recovery of Neisseria gonorrhoeae in clinical specimens. However, there are Culture plates were examined at 24 & 48 hours and bacteria presumptively demonstrates an increase in recovery of the target thereby contributing to numerous disadvantages that make this procedure increasingly less identified on the basis of colony morphology, oxidase (positive), Gram stain a service improvement.
attractive in a busy, cost-driven, clinical environment. Direct inoculation (Gram negative diplococci), and confirmed by biochemical reactions using API® requires parameters which the laboratories are unable to strictly control Growth of Neisseria gonorrhoeae was achieved in a total of 14 samples NH Biomerieux (Product Code:10 400) (Acceptable profile 1000 & 1001) and such as : agar plates have to be purchased, transported to, refrigerated, and obtained from the Sigma Transwab® plates, while only 12 positive cultures Reference laboratory confirmation by Scottish Bacterial Sexually Transmitted kept in-date at Sexual Health Clinics. Due to the large geographical area were isolated from the direct culture plates, a 14% increase in recovery Infections Reference Laboratory, NRIE, Edinburgh.
served by NHS Fife, culture plates inoculated in the Sexual Health Clinics capability. Growth was achieved from the Sigma Transwab® plates at 24 are often delayed in transit to the laboratory which in turn has an impact on hours, in 6 cases, whereas all of the direct culture plates were incubated the recovery of Neisseria gonorrhoeae. The introduction of Sigma for a further 24 hours. The results indicate not only a more rapid Transwab® (manufactured and distributed by Medical Wire) was evaluated identification, but also a more accurate method of diagnosis. This is as a method of improving specimen quality in these circumstances.
demonstrated in the 2 cases where Neisseria gonorrhoea was only detected when using Sigma Transwab®. The current method of direct inoculation would have produced false negative results in these cases.
To determine whether the use of foam-tipped swabs in liquid Amies transport Delays in incubation of direct culture plates along with use of media below medium would result in comparable recovery of Neisseria gonorrhoeae with optimum conditions undoubtedly contribute to the failures of the current direct near patient inoculation of specimens to LCAT culture plates.
system. Management of Sigma Transwab® stock is easier than culture plates as they can be stored at room temperature and have a shelf life of 2 years.
Based on published recovery rate data, literature reviews and user The Sigma Transwab® inoculated plates were stroked out for single preference, the Sigma Transwab® was selected for the evaluation.
colonies which in some cases made the performance of further identification possible at 24 instead of 48 hours. This is particularly evident • MW177S Mini tip swab, orange capped, for urethral sampling.
in rectal and throat swab cultures where mixed cultures are common. • MW176S Standard tipped swab, purple capped, for cervical, rectal, throat and eye sampling.
• LCAT (Lincomycin, Colistin sulphate, Amphotericin B, Trimethoprim)– The use of Sigma Transwab® in liquid Amies transport medium provides a Thermofisher product no: PB0226A). superior recovery method for Neisseria gonorrhoeae from clinical material which is not only a quality improvement but also a service improvement for the patient, allowing for a more efficient management of the process.
The outcome of this study was a change in procedure at all Sexual health The Sigma Transwabs® were obtained in parallel with the direct culture on clinics. NHS Fife adopted the use of Sigma Transwab® in liquid Amies LCAT plates in three Sexual health clinics over a 7 month period. Patients transport medium to obtain the relevant anatomical samples for were informed that the trial swabs would be obtained. If the patient declined transportation to the Microbiology laboratory for culture and identification the request this was noted in the clinical notes, "patient declined trial of Neisseria gonorrhoeae.
Direct culture/ Trial swab POS: Neisseria gonorrhoeae isolated Procedure for direct culture to LCAT agar plates Direct culture/ Trial swab NEG: Neisseria gonorrhoeae NOT isolated Acknowledgements and References
▪ 10µl disposable loops for urethral sampling were used to prepare a smear Thank you to the Sexual Health clinic staff, NHS Fife. Microbiology and Infection control slide and inoculate a single LCAT culture plate. Of a possible 200 samples, 177 trial swabs were obtained from the 3 staff, North Laboratory & K. Graveling for assistance and advise.
▪ Urine sample was obtained for NAAT (Nucleic Acid Amplification Test - selected trial sites. Confirmed Neisseria gonorrhoeae was isolated Roche) for Chlamydia trachomatis and Neisseria gonorrhoeae PCR testing. 1. Olsen, C et al (1999). Comparison of direct inoculation and Copan transport system for from 14 samples. ▪ For all other anatomical sites (cervical, throat, rectal and eye) the NAAT isolation of Neisseria gonorrhoeae from endocervical specimens. J.Clin. Micro. 37:3583-3585.
sample was collected, followed by inoculation of a single LCAT culture plate 2. Thompson, D et al (1999). Comparison of commercial Amies transport systems with using a dry cotton swab for each site.
inhouse Amies medium for recovery of Neisseria gonorrhoeae. J.Clin.Micro: 37:3020-3021.
3. Chernesky, M et al (2006). Use of flocked swabs and a universal transport medium to Procedure for Sigma Transwab® sample collection enhance molecular detection of Chlamydia trachomatis and Neisseria gonorrhoeae. J.Clin.Micro: 44:1084-1086.
4. Public Health England, UK Standards for Microbiology Investigation, B28 Investigation of ▪ Male urethral sampling - the Sigma Transwab® was obtained before the Trial swab not obtained genital tract and associated specimens. B28i4.3 (Dec 2012) urine sample for NAAT. The swab was rolled over visible discharge or Trial swabs taken 5. Public Health England, UK Standards for Microbiology Investigation, ID6 –Identification inserted into but not beyond the terminal 2 cm of the urethra.
of Neisseria species, ID6i2.1 (Sept 2007) ▪ For all other sites the Sigma Transwab® was obtained after all other 6. Medical Wire – wwwTranstube and Sigma Transwab® with liquid modified Amies samples for NAAT and direct culture plate inoculation had taken place. medium –MW176S & MW177S. 1. Biomedical Scientist Microbiology 2. Consultant Microbiology 3. Consultant

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