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Journal of Clinical Microbiology, May 1998, p. 1271-1276, Vol. 36, No. 5
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Typing of Helicobacter pylori vacA Gene and Detection of cagA Gene by PCR and Reverse Hybridization

L. J. van Doorn,1,* C. Figueiredo,2 R. Rossau,3 G. Jannes,3 M. van Asbroeck,3 J. C. Sousa,4 F. Carneiro,2 and W. G. V. Quint1

Delft Diagnostic Laboratory, Delft, The Netherlands1; IPATIMUP2 and Faculty of Pharmacy,4 University of Porto, Porto, Portugal; and Innogenetics N.V., Industriepark Zwijnaarde, Zwijnaarde, Belgium3

Received 10 September 1997/Returned for modification 30 December 1997/Accepted 30 January 1998

The present report describes an analysis of two virulence genes of Helicobacter pylori. Parts of the cagA gene, as well as parts from the signal (s) and middle (m) regions of the mosaic vacA gene, were amplified with biotin-labelled PCR primers and the products were subsequently analyzed by a single-step reverse hybridization line probe assay (LiPA). This assay comprises a strip containing multiple specific probes for the vacA s region (s1a, s1b, and s2 alleles), the vacA m region (m1 and m2 alleles), and the cagA gene. A total of 103 H. pylori-positive materials, including cultured isolates, gastric biopsy specimens, and surgical specimens from patients living in Portugal (n = 55) and The Netherlands (n = 48) were tested by the PCR-LiPA. cagA was detected in 84 and 73% of the Portuguese and Dutch patients, respectively. vacA typing results, as determined by reverse hybridization, were completely concordant with those of sequence analysis. Most Portuguese patients (72%) contained type s1b, whereas most Dutch patients (61%) contained type s1a (P < 0.001). The method is also very effective at detecting the presence of multiple genotypes in a single biopsy specimen. The prevalence of multiple strains in Portuguese patient samples was significantly higher (29%) than that in Dutch patient samples (8%) (P = 0.001). There was a significant association between the presence of ulcers or gastric carcinoma and the presence of vacA type s1 (s1a or s1b; P = 0.008) and cagA (P = 0.003) genes.

* Corresponding author. Mailing address: R. de Graafweg 7, P.O. Box 5010, 2625 AD, Delft, The Netherlands. Phone: 31-15-2604577. Fax: 31-15-2604550. E-mail: L.J.van.Doorn{at}ddl.nl.

Journal of Clinical Microbiology, May 1998, p. 1271-1276, Vol. 36, No. 5
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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