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Clinical and Diagnostic Laboratory Immunology, July 2004, p. 766-769, Vol. 11, No. 4
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.4.766-769.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Novel Hairpin-Shaped Primer Assay To Study the Association of the –44 Single-Nucleotide Polymorphism of the DEFB1 Gene with Early-Onset Periodontal Disease

Michele Boniotto,1 Manzour Hernando Hazbón,2 William James Jordan,1 Greig Patrick Lennon,1 Joyce Eskdale,1 David Alland,2 and Grant Gallagher1*

Department of Oral Biology,1 Division of Infectious Diseases, Department of Medicine, and the Ruy V. Lourenço Center for the Study of Emerging and Reemerging Pathogens, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, New Jersey 07103-27142

Received 20 December 2003/ Returned for modification 17 February 2004/ Accepted 19 March 2004

A powerful, cost-effective new method for studying single-nucleotide polymorphisms (SNPs) is described. This method is based on the use of hairpin-shaped primers (HP), which give a sensitive and specific PCR amplification of each specific allele, without the use of costly fluorophore-labeled probes and any post-PCR manipulation. The amplification is monitored in real-time using SYBR Green I dye and takes only 2 h to yield results. The HP assay has a simple design and utilizes a conventional real-time PCR apparatus. The –44 C->G transversion in the DEFB1 gene (which encodes human ß-defensin 1) has been previously associated with Candida carriage in oral epithelia. In this study, we analyzed the association between early-onset periodontal disease (EOP) and the –44 SNP. We used an HP assay to study the distribution of the –44 SNP in 264 human DNAs obtained from two cohorts of EOP patients and healthy controls from different ethnic backgrounds. The results indicate that the –44 SNP has a similar distribution between EOP and healthy patients, suggesting that it is not associated with the disease.

* Corresponding author. Mailing address: Department of Oral Biology, New Jersey Dental School, 185 South Orange Ave., Newark, NJ 07103. Phone: (973) 972-2971. Fax: (973) 972-2970. E-mail: gallaggr{at}umdnj.edu.

Clinical and Diagnostic Laboratory Immunology, July 2004, p. 766-769, Vol. 11, No. 4
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.4.766-769.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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