Wang Yunxia, Chen Ming, Zhang Liqun, Ding Yi, Luo Yang, Xu Qinghua, Shi Jianfeng, Cao Liang, Fu Weiling
Department of Laboratory Medicine, Southwest Hospital, Third Military Medical University, Chongqing 400038, PR China.
Biosens Bioelectron. 2009 Aug 15;24(12):3455-60. doi: 10.1016/j.bios.2009.04.034. Epub 2009 May 3.
A novel leaky surface acoustic wave (LSAW) bis-peptide nucleic acid (bis-PNA) biosensor with double two-port resonators has been constructed successfully for the quantitative detection of human papilloma virus (HPV). The bis-PNA probe can directly detect HPV genomic DNA without polymerase chain reaction (PCR) amplification, and it can bind to the target DNA sequences more effectively and specifically than a DNA probe. When the concentrations varied from 1 pg/L to 1000 microg/L, with 100 microg/L being the optimal, a typical linearity was found between the quantity of target and the phase shifts. The detection limit was 1.21 pg/L and the clinical specificity was 97.22% of that of real-time PCR. The bis-PNA probe was able to distinguish sequences that differ only in one base. Both the intraassay and interassay coefficients of variance (CVs) were <10%, and the biosensor can be regenerated for ten times without appreciable loss of activity. Therefore, this technical platform of LSAW biosensor can be applied to clinical samples for direct HPV detection.
一种新型的具有双二端口谐振器的漏声表面波（LSAW）双肽核酸（bis-PNA）生物传感器已成功构建，用于定量检测人乳头瘤病毒（HPV）。该双肽核酸探针无需聚合酶链反应（PCR）扩增即可直接检测HPV基因组DNA，并且与DNA探针相比，它能更有效、更特异地结合目标DNA序列。当浓度在1 pg/L至1000 μg/L之间变化时，以100 μg/L为最佳，在目标量与相移之间发现了典型的线性关系。检测限为1.21 pg/L，临床特异性为实时PCR的97.22%。该双肽核酸探针能够区分仅相差一个碱基的序列。批内和批间变异系数（CV）均<10%，并且该生物传感器可以再生十次而活性无明显损失。因此，这种LSAW生物传感器技术平台可应用于临床样本以直接检测HPV。
