การพัฒนาชุดตรวจกรองหาเชื้อ Human papillomavirus type 58 ในผู้ป่วยไทยที่เป็นมะเร็งปากมดลูกด้วยเทคนิค Loop-Mediated isothermal amplification

Abstract

Infection with high-risk human papillomavirus is a major risk factor for development of cervical cancer. In this study we developed LAMP technique for detection of HPV type 58 which is one of most prevalent type in Thailand. Loop-mediated isothermal amplification (LAMP) is a rapid method that can amplify nucleic acid under isothermal condition with high specificity and sensitivity. The reaction was incubated at 63 ◦C for 60 min and the amplified product can be visualized directly by naked eye. Therefore, simple equipment such as a regular water bath is enough to maintain the isothermal conditions. The sensitivity and specificity of this assay was shown to be equivalent to nested PCR that could amplify 103 copies of target DNA. The result of LAMP in clinical sample was found to be consistent with nested PCR. Due to its simplicity, rapidity and high sensitivity, LAMP might be used as the alternative technique for HPV detection in medical diagnostic laboratories.