High-resolution human papillomavirus genotyping by MALDI-TOF mass spectrometry

被引：32

作者：

Hong, Sun Pyo ^{[1
]}

Shin, Soo-Kyung ^{[1
]}

Lee, Eun Hee ^{[2
]}

Kim, Eun Ok ^{[1
]}

Il Ji, Seung ^{[1
]}

Chung, Hyun Jae ^{[1
]}

Park, Sun Nie ^{[3
]}

Yoo, Wangdon ^{[1
]}

Folk, William R. ^{[4
]}

Kim, Soo-Ok ^{[1
]}

机构：

[1] GeneMatrix Inc, R&D Ctr, Yongin 446913, South Korea

[2] Green Cross Reference Lab, Dept Lab Med, Yongin 446913, South Korea

[3] Natl Inst Toxicol Res, Korea Food & Drug Adm, Dept Toxicol Res, Seoul 122704, South Korea

[4] Univ Missouri, Dept Biochem, Columbia, MO 65201 USA

来源：

NATURE PROTOCOLS | 2008年 / 3卷 / 09期

关键词：

D O I：

10.1038/nprot.2008.136

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe a matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS)-based assay for human papillomavirus (HPV) genotyping-the restriction fragment mass polymorphism (RFMP) assay, which is based on mass measurement of genotype-specific oligonucleotide fragments generated by TypeIIS restriction endonuclease cleavage after recognition sites have been introduced by PCR amplification. The use of a TypeIIS restriction enzyme makes the RFMP assay independent of sequence and applicable to a wide variety of HPV genotypes, because these enzymes have cleavage sites at a fixed distance from their recognition sites. After PCR amplification, samples are subjected to restriction enzyme digestion with FokI and BtsCI and desalting using Oasis purification plates, followed by analysis by MALDI-TOF MS. Overall, the protocol is simple, takes similar to 4-4.5 h and can accurately detect and identify at least 74 different HPV genotypes.

引用

页码：1476 / 1484

页数：9

共 43 条

[1]

Abdi F., 2001, NUCLEIC ACIDS RES, V29, P61

[2] Association of genetic variations in CCR5 and its ligand, RANTES with clearance of hepatitis B virus in Korea [J].