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Amplification with molecular beacon primers and reverse line blotting for the detection and typing of human papillomaviruses

Amplification with molecular beacon primers and reverse line blotting for the detection and typing of human papillomaviruses
Amplification with molecular beacon primers and reverse line blotting for the detection and typing of human papillomaviruses
A novel method for the detection and typing of human papillomavirus (HPV) was developed using molecular beacon primers. The method is based on the use of HPV-specific primers containing a hairpin loop structure in which fluorescent donor and quencher groups are held in close proximity such that fluorescence is quenched. Amplification of the target sequence results in the opening of the loop and the resulting fluorescence can be detected on a sequence detector system (SDS) 7700 (Applied Biosystems), as used for TaqMan™ assays. Fluorescent amplicons were identified on the SDS 7700 and then typed by a single hybridisation with specific probes immobilised in lines on a nylon membrane and detected on a fluorescent scanner. This novel beacon primer method compared well with conventional PCR for cervical scrape specimens. The combination of the beacon primer method and reverse line blotting should enable large-scale population studies of HPV infection.
human papilomavirus, line blotting, molecular beacon primer, sequence detector system
0166-0934
29-37
Jordens, J.Z.
515490c8-77d5-4432-9cdb-11f8c152df73
Lanham, S.A.
28fdbbef-e3b6-4fdf-bd0f-4968eeb614d6
Pickett, M.A.
4b358cb6-2f2e-4691-a314-0e80df48eadb
Amarasekara, S.
5d072c04-34f6-42d6-9dc2-6971a0e3f5ca
Abeywickrema, I.
76223e05-2619-4048-adb2-5431db985f30
Watt, Peter J.
8a27f505-e0f4-4663-a19a-ef7f156c8863
Jordens, J.Z.
515490c8-77d5-4432-9cdb-11f8c152df73
Lanham, S.A.
28fdbbef-e3b6-4fdf-bd0f-4968eeb614d6
Pickett, M.A.
4b358cb6-2f2e-4691-a314-0e80df48eadb
Amarasekara, S.
5d072c04-34f6-42d6-9dc2-6971a0e3f5ca
Abeywickrema, I.
76223e05-2619-4048-adb2-5431db985f30
Watt, Peter J.
8a27f505-e0f4-4663-a19a-ef7f156c8863

Jordens, J.Z., Lanham, S.A., Pickett, M.A., Amarasekara, S., Abeywickrema, I. and Watt, Peter J. (2000) Amplification with molecular beacon primers and reverse line blotting for the detection and typing of human papillomaviruses. Journal of Virological Methods, 89, 29-37. (doi:10.1016/S0166-0934(00)00195-6). (PMID:10996637)

Record type: Article

Abstract

A novel method for the detection and typing of human papillomavirus (HPV) was developed using molecular beacon primers. The method is based on the use of HPV-specific primers containing a hairpin loop structure in which fluorescent donor and quencher groups are held in close proximity such that fluorescence is quenched. Amplification of the target sequence results in the opening of the loop and the resulting fluorescence can be detected on a sequence detector system (SDS) 7700 (Applied Biosystems), as used for TaqMan™ assays. Fluorescent amplicons were identified on the SDS 7700 and then typed by a single hybridisation with specific probes immobilised in lines on a nylon membrane and detected on a fluorescent scanner. This novel beacon primer method compared well with conventional PCR for cervical scrape specimens. The combination of the beacon primer method and reverse line blotting should enable large-scale population studies of HPV infection.

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More information

Published date: September 2000
Keywords: human papilomavirus, line blotting, molecular beacon primer, sequence detector system
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 348221
URI: https://eprints.soton.ac.uk/id/eprint/348221
ISSN: 0166-0934
PURE UUID: 2380b899-11f3-4d1c-b18b-67629bdcdf6e
ORCID for S.A. Lanham: ORCID iD orcid.org/0000-0002-4516-264X

Catalogue record

Date deposited: 04 Mar 2013 16:57
Last modified: 17 Jul 2019 21:34

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