JOURNAL

OF

CLINICAL MICROBIOLOGY, Feb. 1991, p. 395-397

Vol. 29, No. 2

0095-1137/91/020395-03$02.00/0 Copyright C 1991, American Society for Microbiology

Comparison of Isolates of Helicobacter pylori and Helicobacter mustelae DONNA R. MORGAN,'* JAMES G. FOX,2 AND ROBERT D. LEUNK' Miami Valley Laboratory, The Procter & Gamble Company, P.O. Box 398707, Cincinnati, Ohio 45239-8707,1 and Division of Comparative Medicine, Massachusetts Institute of Technology, Cambridge, Massachusetts 021392 Received 8 June 1990/Accepted 8 November 1990

On the basis of analysis of protein profiles, isolates of Helicobacter pyloni and Helicobacter mustelae were 50% of HeLa cells in a microtiter well indicated the presence of cytotoxic activity. For hemagglutination assays, bacteria were grown on chocolate agar and serially diluted twofold in phosphatebuffered saline (PBS). A 10-,ul amount was placed into 96-well microtiter plates. Ten microliters of PBS or test inhibitor followed by 20 ,ul of erythrocytes (human type A, 2% in PBS) was added and mixed. The plates were incubated overnight at 4°C and then observed for hemagglutination. Protein profiles of H. pylon and H. mustelae. The protein profiles from three H. pylori isolates were compared with those of H. mustelae isolates from five different ferrets (Fig. 1). All three strains of H. pylori were at least 80% similar, with major proteins of 23, 29, 45, 61, 63, and 66 kDa. All five strains of H. mustelae were at lea5t 85% similar, with major proteins of 23, 24, 27, 31, 33, 44, 54, 64, and 65 kDa. Profiles from H. pylori and H. mustelae were less than 40% similar. Therefore, gastric Helicobacter isolates from humans and ferrets are markedly different. Similarity of protein profiles was determined for isolates from five ferrets (with paired isolates from four ferrets). As observed above, all ferret isolates were similar (Fig. 2). Three ferrets (C, D, and E) were colonized with the same strain (>90% similar). Two ferrets (C and E) were colonized with identical strains (>90% similar) on two occasions. In contrast, two ferrets (B and F) were colonized with different strains (320. In contrast, none of the 20 isolates of H. mustelae tested produced cytotoxin. These data suggest that strains of H. mustelae do not possess one of the potential virulence factors of H. pylori. All strains, of both species, hemagglutinated human type A erythrocytes. Hemagglutination of one strain of H. pylori (11638) was inhibited by n-acetylneuramin-lactose (0.40 mg/ ml) or fetuin (0.125 mg/ml), but not by n-acetylneuraminic acid (0.02 mg/ml), asialofetuin (0.125 mg/ml), bovine serum albumin (1.25 mg/ml), or crude porcine gastric mucin (5 mglml). Hemagglutination by the remaining isolates of H. pylori (60190, 133C, and TX30A) and all strains of H. mustelae (n = 13) was inhibited by fetuin, asialofetuin, or mucin, but not by n-acetylneuramin-lactose, n-acetyl-

Corresponding author. 395

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J. CLIN. MICROBIOL.

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FIG. 1. Comparison of protein profiles of isolates of H. pylori and H. mustelae. (A) SDS-PAGE of total proteins of H. pylori (lanes B to E) and H. mustelae (lanes F to J). Each isolate was obtained from a different human or ferret. Lane A contains the molecular mass standards (in kilodaltons). (B) Dendrogram of relatedness among strains of H. pylori and H. mustelae, based on protein profiles. The letters correspond to the lanes in panel A. Strain 11638 (lane D in panel A) was used as a reference strain against which other strains were compared. Therefore, strain 11638 does not appear in the dendrogram.

neuraminic acid, or bovine serum albumin. These data suggest that some strains of H. pylori and H. mustelae share a hemagglutinin. In earlier studies, we used sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) techniques to measure the relatedness of strains of H. pylori. We determined that identical strains of H. pylori were >90% similar and that all strains of H. pylori were >80% similar (13). In

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the current study, we used the same techniques to measure the relatedness of strains of H. pylori and H. mustelae. SDS-PAGE of total protein profiles clearly distinguished H. pylori from H. mustelae. The two species were

Comparison of isolates of Helicobacter pylori and Helicobacter mustelae.

On the basis of analysis of protein profiles, isolates of Helicobacter pylori and Helicobacter mustelae were less than 40% similar. Cytotoxin produced...
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