STREPTOCOCCAL GROUPING

LATEX TEST KIT

Catalogue No Product description Method
STE/010 6x50 Test Kit 1. Using a sterile bacteriological loop, pick 2-6 colonies of streptococci
(avoiding other types of colony on the plate) and emulsify them in 0.4 ml
INTENDED USE extraction enzyme. (If a broth culture is to be grouped, pipette 0.1 ml of
This kit is for the identification of streptococci of Lancefield’s groups A, B, C, D, an overnight culture into 0.4 ml extraction enzyme).
F and G by agglutination of specific antibody - coated latex particles in the 2. Incubate the mixture in a water bath at 37?C for 10 minutes. Shake the tubes
presence of enzymically extracted antigen. vigorously after 5 minutes incubation.
3. Re-suspend the latex reagents by gentle agitation. Dispense 1 drop of each
WARNINGS AND PRECAUTIONS latex onto acircle on the test slide.
For in vitro diagnostic use only 4. Add one drop of the extract from a Pasteur pipette (or another device
For professional use only delivering approximately 50 microlitres) to each drop of latex reagent, and
Health and Safety warnings: mix the contents of each circle with a separate mixing stick.
All patient samples and isolates derived from patient samples and reagents should be 5. Rock the slide for not longer than 1 minute, then observe for agglutination.
treated as potentially infectious and the user should wear protective gloves, eye
protection and laboratory coats when performing the test. Note: The positive control is supplied so that the reactivity of all the latex reagents
Non disposable apparatus must be sterilised after use by an appropriate method. can be checked with each batch of tests. It requires no extraction or dilution before
Disposable apparatus must be treated as biohazardous waste and autoclaved or use, and should be used as in steps 3 to 5 above. All the latex reagents should show
incinerated. strong agglutination within 1 minute.
Spillages of potentially infectious material should be absorbed and disposed of as
above. The site of spillage must be sterilised with disinfectant or 70% alcohol. RESULTS
Do not pipette by mouth. A Positive Result is indicated by the visible agglutination of the latex particles.
These reagents contain micro-fine latex suspensions coated with rabbit serum. The This will normally occur within a few seconds of mixing, depending on the
product also contains aqueous buffer salts including less than 0.1% sodium azide as strength of the antigen extract.
a preservative - see material safety data sheet The dropper bottle teat is made from
natural rubber. A Negative Result is indicated by a milky appearance without any visible
Analytical precautions: agglutination of the latex particles.
Do not modify the test procedure. However, faint traces of granularity may be detected in negative patterns,
Do not dilute or modify the reagent in any way. depending on the visual acuity of the operator.
Allow all reagents and samples to reach room temperature (18 - 30ºC) before use.
Resuspend latex reagent preparation gently but thoroughly. INTERPRETATION OF RESULTS
Discard the reagent if the suspension becomes rough (i.e. shows signs of auto- Strong agglutination with the FIRST latex reagent indicates a positive
agglutination) or fails to agglutinate with cultures known to contain clumping factor identification of that group. Only strong agglutination is significant; occasional
or Protein A. strains of streptococci may give weak reactions with more than one group. Weak
and granular reactions should be ignored. If agglutination occurs in all groups,
COMPOSITION either the enzyme has been over-inoculated in which case repeat the test using a
Kit presentation lighter inoculum, or a mixed culture was tested, in which case check for purity and
1. 6x50T latex determinations for the grouping of streptococci A:B:C:D:F:G. retest. False positive results can occur if the test is continued for longer than one
(Yellow labels) minute.
2. Polyvalent positive control 2ml. (Red label) False positive reactions have been known to occur with organisms from unrelated
3. Freeze Dried Extraction Enzyme. 2 vials. (Green label). Reconstitute each genera, eg. Escherichia, Klebsiella or Pseudomonas. These are likely to non-
with 10ml of distilled water. specifically agglutinate all latex reagents.
4. Disposable test cards x 50. The group D antigen is common to organisms of groups Q,R and S.
5. Mixing sticks 300 and kit insert. False negative results can occur if an inadequate amount of culture is used for
extraction.
STORAGE AND SHELF LIFE
Store latex reagents and controls upright at 2-8ºC. PERFORMANCE CHARACTERISTICS AND LIMITATIONS OF
DO NOT FREEZE LATEX REAGENTS. THE METHOD
Do not use reagents after the stated expiry date.
Once opened latex reagents may be used until the expiry date provided they have Plasmatec
been stored correctly and have not been contaminated. + -
The freeze dried Extraction Enzyme should be stored at 2-8? C. Once reconstituted
with 10ml of sterile distilled water, it will retain its activity for at least 3 months or
Reference

+ 607 55
method

until the date shown on the bottle label, whichever is sooner. Alternatively the
enzyme may be stored in aliquots of 0.4ml frozen at -20? C, when it will remain
active for at least 6 months or until the date shown on the original bottle, whichever
is the sooner. - 0 24
Do not freeze and thaw more than once!

MATERIALS AND EQUIPMENT REQUIRED BUT NOT PROVIDED Sensitivity 607/662 = 92%
Water bath @ 37? C. Specificity 24/24 = 100%
Test tubes.
Pasteur and graduated pipettes.
INTERNAL QUALITY CONTROL
A positive control is provided and should be used to verify that the latex reagents are
SPECIMEN AND SAMPLE PREPARATION working satisfactorily under test conditions.
Cultures
Periodically check the following:
Note colonial characteristics, haemolysis, and cell morphology before starting the
1. The test reagents agglutinate with a known reference Streptococcus strain
test. Ensure that the organisms to be tested are Gram-positive and catalase-
2. The test reagents do not auto agglutinate in normal saline solution.
negative. Any blood agar plate culture yielding 2-6 well- separated colonies may
be used, they should have been inoculated from a pure culture of the organism.
REFERENCES
1. Lancefield, R.c., (1938)Proc. Soc.Exp. Bio. Med. 38, 473
PROCEDURE 2. Harvey,C.L.,Mcillmurray, M.B. (1984) Eur.J. Clin. Microbiol, 3.6,526
Principle
3. Facklam,R.R., (1980) “Manual of Clinical Microbiology” 3rd Edn., American
Streptococci carry group specific carbohydrate antigens in their cell walls. After
Society for Microbiology, Washington, DC, pp 88-110.
extraction by a specially developed enzyme preparation these antigens will
agglutinate latex particles coated with the corresponding antibody. The latex
remains in smooth suspension in the absence of group specific antigen.
PSTE.V3 4/4/03