Overview: Detection of Human Papillomavirus in Clinical Samples.

Human papillomavirus (HPV) is the most common sexually-transmitted virus and it is known that persistent infection by high-risk HPV is a necessary factor for cervical carcinogenesis. Although cytological screening has decreased the incidence of cervical cancer, the sensitivity and specificity of testing is limited. To date, HPV-driven molecular techniques have provided a number of potential biomarkers for both diagnostic and prognostic use in clinical management. In addition, they can provide insights into the biology of HPV-induced cancers leading to non-surgical therapy. This review summarizes current knowledge of detection methods for HPV and related biomarkers that can be used to discriminate lesions with a high risk of progression of cervical cancer.

The evaluation of loop-mediated isothermal amplification-quartz crystal microbalance (LAMP-QCM) biosensor as a real-time measurement of HPV16 DNA.

We have previously developed quartz crystal microbalance biosensor integrated with loop-mediated isothermal amplification (LAMP-QCM) for human papillomavirus (HPV) type58 DNA detection. Infection with HPV, particularly HPV16, remains a serious health problem due to its major risk factor contributing to cervical cancer. In the present study, LAMP-QCM biosensor was evaluated in terms of a quantitative assay for copy number of HPV16 DNA in cervical samples compared to quantitative PCR using TaqMan assay (TaqMan-qPCR). The detection limit of LAMP-QCM was found to be 10 fold more sensitive than TaqMan-qPCR with 100% specificity and 7.6% imprecision. Different plot of HPV16 DNA copy number using Bland-Altman analysis revealed 94% correlation between LAMP-QCM and qPCR. We therefore concluded that the developed LAMP-QCM biosensor provides a possible rapid and sensitive assay for HPV16 DNA quantification in a routine laboratory.

Quartz crystal microbalance biosensors: prospects for point-of-care diagnostics.

BACKGROUND: A QCM is a label-free and extremely mass-sensitive device, which allows the detection of the binding event between trace medical analytes and bio-receptors on its surface. QCM, the most promising type of biosensors, has attracted much interest due to the inherent benefits over other transducers, including better sensitivity, ease-of-use, integration with compact analytical devices, and economy, and also involving relatively simple technology in its production. Thus, they have great potential with regard to point-of-care (POC) testing for early detection of diseases. MATERIAL AND METHOD: Retrievable articles that related to acoustic type sensing of Pubmed and Science direct database were included. Additionally, abstracts presented at Biosensor World Congress held between 2008 and 2012 were searching to identify relevant clinical trials. RESULTS: All studies demonstrated the opportunity in the use of QCM as a novel diagnostic method. Several attempts have been made to construct integrated systems that show promising application for POC tests. CONCLUSION: This review represents another step to meet challenges, especially in the improved minimization and sensitivity of biosensors. As this work continues, new bioreceptor and biomarkers emerging from the could make it an ideal candidate for cheap POC diagnostic.

The development of DNA-based quartz crystal microbalance integrated with isothermal DNA amplification system for human papillomavirus type 58 detection.

To address the effect of dramatic change in temperature and viscosity during PCR process on quartz crystal microbalance (QCM) sensor and to increase the sensitivity, isothermal amplification was employed in the system. We combined loop-mediated isothermal amplification (LAMP) technique with QCM, called as LAMP-QCM, for detection of high-risk human papillomavirus viral DNA type 58 (HPV-58) which is commonly found in Asian women. The liquid-phase LAMP-QCM prototype comprised the frequency counter, a temperature control device and housing of the quartz crystal with polished gold electrodes on both sides. QCM detection signal was monitored in real-time based on an avidin-biotin binding between avidin coated QCM surface and specific biotinylated LAMP products. Analytical performance was evaluated for precision, sensitivity and specificity. A plasmid clone containing the HPV-58 sequence was diluted from 10(6) to 1 copy and used for detection limit. Cut-off value was estimated at 28.8 Hz from negative viral template. The system could detect 100 copies with Δf at 34.0±3.6 Hz compared to 1000 copies detected by conventional LAMP. No cross-reaction was observed with other HPV types. The HPV-58 detection was compared among LAMP-QCM, conventional LAMP and nested PCR in 50 cervical cancer tissues. The positive rate of LAMP-QCM was higher than that of conventional LAMP with 100% sensitivity and 90.5% specificity. The integrated LAMP-QCM system has improved the detection limit up to ten times compared to conventional LAMP with less-time consuming.