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Journal of Clinical Microbiology, October 2008, p. 3546-3547, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.01027-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

LETTER TO THE EDITOR

Evaluation of Three Commercial Latex Agglutination Tests for Identification of Campylobacter spp.{triangledown}


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LETTER
 
Latex agglutination tests for the rapid identification of Campylobacter spp. have been available in the market for more than 20 years (1, 3). As of November 2007, there were only three commercial latex agglutination tests: PanBio-Campy (jcl) (PanBio Inc., Columbia, MD), the Dryspot Campylobacter (Oxoid, Basingstoke, Hampshire, England), and the Microgen M46 Campylobacter (Microgen Bioproducts Ltd., Camberley, Surrey, United Kingdom). The only test approved by the Food and Drug Administration of the United States is PanBio-Campy (jcl), which was approved under the name Meritec-Campy (jcl) by Meridian Biosciences, Inc. (Cincinnati, OH; formerly known as Meridian Diagnostics), in 1993. However, as of August 2008, PanBio-Campy will be produced and marketed by Scimedx Corporation (Denville, NJ) under the name CAMPY (jcl).

These latex agglutination tests use polyclonal antibodies to detect antigenic outer membrane proteins or antigenic epitopes from flagella. We evaluated these three test kits in parallel with a variety of Campylobacter strains belonging to eight species. These strains have been isolated from clinical and food sources. We also tested these kits with strains of bacteria other than Campylobacter, mostly isolated from clinical and food sources. These strains included six Acinetobacter baumannii strains and one strain each of Arcobacter butzleri, Arcobacter skirrowii, Listeria monocytogenes, Salmonella enterica subsp. enterica, Salmonella sp., and Shigella sp. These strains were selected based on phylogenetic similarities to Campylobacter (Arcobacter) spp. and potential growth as contaminants on plate media used for isolation of Campylobacter (Acinetobacter) spp. (5) and for comparison with other food-borne pathogens (Listeria monocytogenes). In December 2007, a new product (Campylobacter Latex test, reference no. 96143) was released to the market by Liofilchem Immunology, SRL (Italy), for the identification of Campylobacter jejuni. This test was not included in the evaluation. We are not aware of any microbiology company distributing this recent test from Italy in the United States.

Among the Campylobacter strains, those numbered *0.07 or *0.08 were isolated from pediatric enteritis patients at the Groote Schuur Hospital, Cape Town, South Africa, in 2007 and 2008, as indicated in Table 1. These isolates were identified to the species level by the protocol outlined by Lastovica (2). The rest of the isolates belong to the culture collection of the Microbiology Laboratory, Department of Poultry Science, Auburn University. These strains are ATCC (American Type Culture Collection) strains or were isolated from food samples at Auburn University from 2005 to 2007. C. jejuni and C. coli strains have been identified with two multiplex PCR assays described elsewhere (4).


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  		TABLE 1. Strains of Campylobacter species used in these studies

Table 1 shows the results of testing 125 Campylobacter strains with the three latex tests. The Microgen Campylobacter M46 test reacted with 123 strains of all eight Campylobacter spp. This test did not react with one C. jejuni (1266) and one C. upsaliensis (ATCC 43953) strain. Microgen Bioproducts specifies that the M46 Campylobacter test reacts with thermotolerant species such as C. jejuni, C. coli, C. lari, and C. upsaliensis and emerging Campylobacter spp. such as C. jejuni subsp. doylei, C. concisus, and C. hyointestinalis. The Oxoid test reacted with 105 strains of six Campylobacter spp. This test failed to react with one C. jejuni strain (1266) and one C. upsaliensis strain (ATCC 43953), the same strains with which the Microgen test did not react. According to Oxoid, this test reacts only with thermotolerant species (C. jejuni, C. coli, C. lari, and C. upsaliensis). As stated by the manufacturer, the PanBio test reacted only with C. jejuni, C. coli, and C. lari. However, this test did not react with two C. coli strains (ATCC BAA-371 and 1315).

The Oxoid test was the only one that did not react with any of the non-Campylobacter bacterial strains tested (Table 2). The Microgen test reacted with an A. baumannii and a Listeria monocytogenes strain, but the reactions were in the form of large clumps and could easily be differentiated from the regular, fine agglutination seen with positive samples. The PanBio test reacted with two A. baumannii strains, and although one reaction was also in the form of large clumps, one strain yielded a very fine agglutination that was difficult to differentiate from the agglutination produced by a positive test.


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  		TABLE 2. Non-Campylobacter species tested in these studies

All three tests were simple and easy to use and had a reasonable shelf life when stored as directed by the manufacturer. Of the three tests, the most expensive is the Oxoid test, while the PanBio test is the least expensive. For the analysis of food samples, where C. jejuni and C. coli are the main suspected species and where large numbers of samples are tested, the PanBio test may be an economical option. However, cross-reactivity was evident with an A. baumannii strain, a common organism growing on agar plates used for Campylobacter isolation (5). Therefore, precise identification of Campylobacter isolates requires the use of phase-contrast microscopy or Gram staining to avoid misidentifications.

Our results indicate that the Microgen Campylobacter M46 test is the most appropriate for the testing of any Campylobacter isolates collected from human and food samples, even for Campylobacter spp. other than C. jejuni and C. coli.


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FOOTNOTES
 
{triangledown} Published ahead of print on 27 August 2008. Back


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REFERENCES
 
    1
  1. Hodinka, R. L., and P. H. Gilligan. 1988. Evaluation of the Campyslide agglutination test for confirmatory identification of selected Campylobacter species. J. Clin. Microbiol. 26:47-49.[Abstract/Free Full Text]
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  2. Lastovica, A. J. 2006. Emerging Campylobacter spp.: the tip of the iceberg. Clin. Microbiol. Newsl. 28:49-56.[CrossRef]
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  3. Nachamkin, I., and S. Barbagallo. 1990. Culture confirmation of Campylobacter spp. by latex agglutination. J. Clin. Microbiol. 28:817-818.[Abstract/Free Full Text]
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  4. Oyarzabal, O. A., S. Backert, M. Nagaraj, R. S. Miller, S. K. Hussain, and E. A. Oyarzabal. 2007. Efficacy of supplemented buffered peptone water for the isolation of Campylobacter jejuni and C. coli from broiler retail products. J. Microbiol. Methods 69:129-136.[CrossRef][Medline]
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  5. Oyarzabal, O. A., K. S. Macklin, J. M. Barbaree, and R. S. Miller. 2005. Evaluation of agar plates for direct enumeration of Campylobacter spp. from poultry carcass rinses. Appl. Environ. Microbiol. 71:3351-3354.[Abstract/Free Full Text]
Robert S. Miller
Leslie Speegle
Omar A. Oyarzabal*
Department of Poultry Science
260 Lem Morrison Dr.
Auburn University
Auburn, Alabama 36849

Albert J. Lastovica
Department of Biotechnology
Faculty of Natural Sciences
University of the Western Cape
Bellville, South Africa

* Phone: (334) 844-2608 Fax: (334) 844-2641 E-mail: oyarzoa{at}auburn.edu

Journal of Clinical Microbiology, October 2008, p. 3546-3547, Vol. 46, No. 10
0095-1137/08/$08.00+0     doi:10.1128/JCM.01027-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.




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