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STAPHYTECT PLUS

Code: DR0850

Staphytect Plus is a latex slide agglutination test 1 for the differentiation of Staphylococcus aureus by detection of clumping factor, Protein A and certain polysaccharides found in methicillin resistant Staphylococcus aureus (MRSA) from those Staphylococci that do not possess these properties.

Introduction
Traditionally, differentiation between coagulase-positive and coagulase-negative staphylococci has been performed either with the tube coagulase test that detects extracellular staphylocoagulase or the slide coagulase test that detects the clumping factor (bound coagulase) present on the bacterial cell surface. Several other differentiation tests are also available including the passive haemagglutination test (Oxoid Staphylase DR0595) and the DNase test.
It has been reported that approximately 97% of human strains of Staphylococcus aureus possess both bound coagulase and extracelluar staphylocoagulase.
Protein A is found on the cell surface of about 95% of human strains of Staphylococcus aureus and has the ability to bind the Fc portion of immunoglobulin G (IgG) 2 .
It has been observed that certain methicillin-resistant strains of Staphylococcus aureus (MRSA) may express undetectable levels of clumping factor and Protein A3,4,5 . It has been shown however that these strains all possess capsular polysaccharide 6. The capsule can mask both Protein A and the clumping factor thereby preventing agglutination.
Staphytect Plus uses blue latex particles coated with porcine fibrinogen and rabbit IgG including specific polyclonal antibodies raised against capsular polysaccharides of Staphylococcus aureus 7,8.
When the reagent is mixed on a card with colonies of Staphylococcus aureus, rapid agglutination occurs through the reaction between (i) fibrinogen and clumping factor, (ii) Fc portion of IgG and Protein A (iii) specific IgG and capsular polysaccharide. Agglutination may also occur with other species which possess clumping factor or Protein A such as Staphylococcus hyicus and Staphylococcus intermedius. If neither clumping factor, Protein A or specific capsular polysaccharides are present, agglutination will not occur and the result will be regarded as negative. The most frequent coagulase and Protein A negative isolates of staphylococci are Staphylococcus epidermidis.

Components of the Kit
DR0851
Staphytect Plus Test Reagent (5.6 ml)
Blue latex particles coated with both porcine, fibrinogen and rabbit IgG together with specific polyclonal antibodies raised against capsular polysaccharide of Staphylococcus aureus. Each bottle contains sufficient reagent for 100 tests.
DR0852 Staphytect Plus Control Reagent (5.6 ml)
Blue unsensitised latex particles. Each bottle contains sufficient reagents for 100 tests.
DR0500 Reaction Cards
There are 35 disposable reaction cards provided in the kit
Instruction Leaflet

Materials required but not provided:
Timer
Microbiological Loop
A suitable laboratory disinfectant
Positive Control: Staphylococcus aureus strain such as ATCC®25923.
Negative Control: Staphylococcus epidermidis strain such as ATCC®12228.

Storage
This kit must be stored at 2-8°C away from direct sunlight or heat sources. Do not Freeze.
The kit should not be used after the expiry date printed on the outside of the carton.

Precautions
Reagents contain 0.095% sodium azide as a preservative. Sodium azide is toxic and may react with lead or copper plumbing to produce metal azides which are explosive by contact detonation. To prevent azide accumulation in plumbing flush with copious amounts of water immediately after waste disposal.
Specimen materials may contain pathogenic organisms handle with appropriate precautions.

References:
1. Essers, L. and Radebold, K. (1980). "Rapid and Reliable Identification of Staphylococcus aureus by a Latex Agglutination Test". J.Clin.Microbiol. 12: 641-643.
2. Taussig, M.J. (1984). Processes in Pathology and Microbiology 2nd Ed. 520-530. Blackwell,Oxford.
3. Ruane, P.J., Morgan, M.A., Citron, D.M. and Mulligan, M.E. (1986). "Failure of Rapid Agglutination Methods to Detect Oxacillin-Resistant Staphylococcus aureus". J.Clin.Microbiol. 24: 490-492.
4. Roberts, J.I.S. and Gaston, M.A. (1987). "Protein A and coagulase expression in epidemic and non-epidemic Staphylococcus aureus". J.Clin.Pathol. 40: 837-840.
5. Wanger, A.R., Morris, S.L. Ericsson, C., Singh, K.V. and LaRocco, M.T. (1992). "Latex Agglutination-Negative Methicillin-Resistant Staphylococcus aureus Recovered from Neonates: Epidemiologic Features and Comparison of Typing Methods". J.Clin. Microbiol. 30: 2583-2588.
6. Fournier, J.M., Boutonnier, A. and Bouvet, A. (1989). "Staphylococcus aureus Strains Which Are Not Identified by Rapid Agglutination Methods Are of Capsular Serotype 5". J.Clin.Microbiol. 27: 1372-1374.
7. Fournier, J.M., Bouvet, A. Boutonnier, A., Audurier, A. , Goldstein, F., Pierre, J., Bure, A., Lebrun, and L., Hochkeppel, H.K. (1987). Predominance of Capsular Polysaccharide Type 5 among Oxacillin-Resistant Staphylococcus aureus". J.Clin.Microbiol. 25: 1932-1933.
8. Karakawa, W.W., Fournier, J.M., Vann, W.F., Arbeit, R., Schneerson, R.S., and Robbins, J.B. (1985). "Method for the Serological Typing of the Capsular Polysaccharides of Staphylococcus aureus ". J.Clin.Microbiol. 22: 445-447.
9. Kloos, W.E., Jorgensen J.H.(1988). Staphylococci. pp.143-153. In Manual of Clinical Microbiology. 4th Edn. (Eds) Lennette, E.H., Balows, A.Hauser W.J., and Shadomy, H.J. Assoc.Amer.Microbiol. Washington.
10. Data on file at Oxoid Ltd.
11. Jean-Pierre, H., Darbas, H., Jean-Roussenq,. A. & Boyer, G.(1989). "Pathogenicity in Two Cases of Staphylococcus schleiferi, a Recently Described Species". J.Clin.Microbiol.27: 2110-2111.
12. Frenney, J., Brun, Y., Bes, M., Meugnier H., Grimont, F., Grimont, P.A.D.,Nervie, C., & Fleurette, J. (1988). "Staphylococcus schleiferi sp. nov. Two species from Human Clinical Specimens". Int.J.Sup Bacteriol. 38: 168-172.
13. Phillips, W.E., and Kloos, W.E. (1981). "Identification of Coagulase-Positive Staphylococcus intermedius and Staphylococcus hyicus. subsp. hyicus Isolates from Veterinary Clinical Specimens". Clin.Microbiol:14: 671-673.
14. van Griethuysen, A., Bes, M., Etienne, J., Zbinden, R. and Kluytmans, J. (2000). "An International Multicenter Evaluation of a new Latex Agglutination Test for Identification of Staphylococcus aureus". Clin Microbiol. and Infect. 6 sup 1: 163.
15. Schnitzler, N., Rainer, M., Conrads, G., Frank, D. and Haase, G. (1988). “ Staphylococcus lugdunensis: Report of a case of Peritonitis and an Easy-To-
Perform Screening Strategy”. J. Clin.Microbiol. 26: 1939–1949.
16. Ann-Herbert, A., Crowder, C. G., Hancock, G. A., Jarvis, W. R. and Thornsberry, C. (1998). “Characteristics of Coagulase-Negative-Staphylococci
That Help Differentiate These Species of the Family Micrococcaceae”. J. Clin.Microbiol. 36: 812–813.
17. Gregson, D. B., Low, D. E., Skulnick, M.and Simor, A. E. (1988). “Problems with Rapid Agglutination Methods for Identification of Staphylococcus aureus
When Staphylococcus saprophyticus Is Being Tested”. J. Clin. Microbiol. 26:1398–1399.
18. Myhre, E. B. and Kuusela, P. (1983).“ Binding of Human Fibronectin to Group A, C and G Streptococci”.Infect. Immun. 40: 29–34.
19. Runehagen, A., Schonbeck, C., Hedneru, Hessel, B. and Kronvall, G.(1981). “Binding of Fibrinogen Degradation Products to S. aureus and to ß-Hemolytic Streptococci Group A, C and G”. Acta. path. microbiol. Scand., Sect B. 89: 49–55.

 
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