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Journal of Clinical Microbiology, December 2001, p. 4575-4576, Vol. 39, No. 12
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.12.4575-4576.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Improved Rate of Isolation of Neisseria
meningitidis by Direct Plating of Pharyngeal
Swabs
Richard
Cunningham,1,*
Roger
Matthews,1
Gill
Lewendon,2
Sarah
Harrison,2 and
James
M.
Stuart3
Plymouth Public Health Laboratory, Derriford
Hospital, Plymouth,1 South and West
Devon Health Authority, Dartington,2 and
Communicable Disease Surveillance Centre (South West),
Public Health Laboratory, Gloucester,3 United
Kingdom
Received 7 June 2001/Returned for modification 26 August
2001/Accepted 15 September 2001
 |
ABSTRACT |
Culturing of pharyngeal swabs for Neisseria
meningitidis is an important clinical and epidemiological tool.
Routine methods include direct plating onto solid medium or later
plating in the laboratory. A comparison of these methods used with 490 high school students found a significantly higher carriage rate with
direct plating (11.8 versus 6.1%; P < 0.001).
 |
TEXT |
Pharyngeal carriage of
Neisseria meningitidis is common in the general population,
with rates of 20 to 30% in young adults (2, 3). Immediate
plating of cotton swabs onto solid culture medium at the site of
swabbing (direct plating) is commonly used in epidemiological studies
to measure carriage rates (2). An accepted alternative
method is to place the swabs in transport medium and send them to the
laboratory for plating (3). In clinical practice in the
United Kingdom, laboratory plating is usual, as reflected in the Public
Health Laboratory Service standard operating procedure for culture of
pharyngeal swabs. The assumption is that prompt (i.e., often overnight)
transport under appropriate conditions will not significantly affect
the rate of isolation of N. meningitidis.
Nasopharyngeal swabs were collected in autumn 1999 from sixth-form
students in the Plymouth area of southwest England as part of a United
Kingdom multicenter study of meningococcal carriage. Other study
centers were in Manchester, Oxford, Nottingham, London, Glasgow,
Cardiff, and Bangor. The study protocol allowed either direct plating
or transport in Amies medium for laboratory plating, with the latter
method being used in Plymouth. The estimated interval between specimen
collection and inoculation of plates ranged between 5 and 7 h. An
interim analysis, after swabs were taken from 906 students, showed an
unexpectedly low meningococcal carriage rate of 6.1% (55 of 906). This
contrasted sharply with results from other centers, where direct
plating was mostly used and where carriage rates ranged from 10 to
23%. The culture medium used was GCVCAT. This is a modified New York
City medium, containing vancomycin, trimethoprim, and amphotericin B. Quality control of the medium revealed no inhibition of meningococcal
growth, and the swab collection technique was satisfactory.
We set out to compare meningococcal isolation rates obtained with
direct and laboratory plating. Pharyngeal swabs from 490 students were
plated on-site and then placed in transport medium for plating at the
laboratory. The carriage rate from direct plating was 11.8% (58 of
490), whereas the rate obtained from laboratory plating was 6.1% (30 of 490) (Table 1). This difference was
highly statistically significant using McNemar's test
(P < 0.001). The carriage rate from laboratory plating
during the comparative study (6.1%) was the same as the rate before
the study (6.1%), suggesting that the difference was unlikely
to be due to loss of organisms from the plate. Swabs were taken from
students in many of the same schools for direct plating a year later.
For schools in which swabs were laboratory plated in the first year and
directly plated in the second year, carriage rates increased from 50 of
761 (6.6%) to 161 of 1,076 (14.9%) (P < 0.0001). For
schools in which swabs were directly plated in both years, carriage
rates were unchanged, i.e., 42 of 342 (12.2%) versus 61 of 504 (12.1%). These data provide further evidence of higher yields from
direct plating.
Detection of meningococcal carriage is important in epidemiological
studies of prevalence, acquisition, and transmission (2, 3,
5) and possibly in outbreak management. Swabbing is also important in individual patient management, as the yield of positive cultures from pharyngeal swabs, as opposed to blood, may remain high
for some time after administration of antibiotics (1). The
sensitivity of a single swab may be low (5, 6), so any means of maximizing yield is important. The sensitivity of any bacterial culture is greatest when the specimen is inoculated directly
onto culture medium immediately after collection. This effect may be
particularly important when attempting culture of Neisseria
species, as has been reported previously with Neisseria gonorrhoeae (4, 7). The benefits of immediate
inoculation must be weighed against the practical difficulties of
providing sterile culture medium to the site of specimen collection. A
study with military personnel found no clear difference between
carriage rates from direct plating and laboratory plating after 5 h in transport medium, but the numbers in that study were too low to exclude such an effect (5). The experience at the
Manchester Public Health Laboratory in two separate carriage studies
with teenagers gave similar rates (21 to 23%) despite direct plating in one study and laboratory plating (mean delay, 3.5 h) in the other. Direct comparison as performed in Plymouth, however, has not
been undertaken (E. Kaczmarski, personal communication).
The reasons for the higher rates of overall carriage found in the
Manchester studies are unexplained and deserve further investigation. Nonetheless, our findings indicate that putting swabs in transport medium for more than 5 h as an alternative to direct plating may halve the culture positivity rate. We recommend direct plating of
pharyngeal swabs whenever practical, both for studies of meningococcal carriage and for investigation of cases of suspected meningococcal disease.
 |
ACKNOWLEDGMENTS |
We are grateful for comments by Ed Kaczmarski.
This work was carried out as a part of the United Kingdom Meningococcal
Carriage Group study into carriage of hypervirulent meningococci before
and after the introduction of serogroup C conjugate polysaccharide
vaccine in the United Kingdom. This multicenter study is funded by the
Wellcome Trust, the National Meningitis Trust, and the Chief Scientist
Office of the Scottish Executive Health Department.
 |
FOOTNOTES |
*
Corresponding author. Mailing address: Plymouth Public
Health Laboratory, Derriford Hospital, Plymouth PL6 8DH, United
Kingdom. Phone: 44 (0) 1752 792387. Fax: 44 (0) 1752 771561. E-mail:
richard.cunningham{at}phnt.swest.nhs.uk.
 |
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Journal of Clinical Microbiology, December 2001, p. 4575-4576, Vol. 39, No. 12
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.12.4575-4576.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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