DNA Modulates the Interaction of Genetically Engineered DNA-Binding Proteins and Gold Nanoparticles: Diagnosis of High-Risk HPV Infection

Abstract

Gene detection has an important role in diagnosing several serious diseases and genetic defects in modern clinical medicine. Herein, we report a fast and convenient gene detection method based on the modulation of the interaction between a heat-resistant double-stranded DNA (dsDNA)-binding protein (Sso7d) and gold nanoparticles (Au NPs). We prepared a recombinant Cys-Sso7d, which is Sso7d with an extra cysteine (Cys) residue in the N-terminus, through protein engineering to control the interaction between Sso7d and Au NPs. Cys-Sso7d exhibited a stronger affinity for Au NPs and more easily induced the aggregation of Au NPs than Sso7d. In addition, Cys-Sso7d retained its ability to bind with dsDNA. The aggregation of Au NPs induced by Cys-Sso7d was diminished in the presence of dsDNA, which could be utilized as a transduction mechanism for the detection of the polymerase chain reaction (PCR) products of human papillomavirus (HPV) gene fragments (HPV types 16 and 18). The Cys-Sso7d/Au NP probe could detect as few as 1 copy of the HPV gene. The sensitivity and specificity of the Cys-Sso7d/Au NP probe for Pap smear clinical specimens (n = 52) for HPV 16 and HPV 18 detection were 85.7%/100.0% and 85.7%/91.7%, respectively. Our results demonstrate that the Cys-Sso7d/Au NP probe can be used to diagnose high-risk HPV types in Pap smear samples with high sensitivity, specificity, and accuracy.

Nanotechnology-Based Weapons to Combat Human Papillomavirus Infection Associated Diseases

“…For example, Sso7d-modified Au NPs have been constructed by Ju-Yi Mao et al to detect HPV genes with a detection limit of 1 copy of gene in specimens. The sensitivity and specificity data of this special probe from clinical specimens of Pap smear ( n = 52) are exciting, as high as 85.7%/100% and 85.7%/91.7%, respectively ( Mao et al, 2017 ). In addition to the application of the AuNPs, alone or combined with other techniques, a hybridization assay composed of quantum dots and superparamagnetic nanoparticle in detecting HPV 16 infections, or the mass spectrometry genotyping of HPV based on high-efficiency selective enrichment of nanoparticles, are both proved to be feasible than traditional methods ( Yu-Hong et al, 2011 ; Zhu et al, 2018b ).…”

Advancements and Future Predictions on Diagnostic Approaches towards Cervical Cancer through Nanotechnology-Based Sensors for the Detection of Human Papillomavirus

“…In this DNA analysis the label-free Cys-Sso7d/Au NP probe as compared to other nanoparticle-based biosensors does not need modified complementary probe. Thus, providing great speed, sensitivity and specificity with cost effectiveness, this assay helped in the detection of high-risk HPV and to improve the accuracy of cervical cancer screening [52]. EIS of the GC-GNS modified electrode was compared with GE.…”

Development of label-free gold nanoparticle based rapid colorimetric assay for clinical/point-of-care screening of cervical cancer

“…The Cys-Sso7d/Au NP probe coupled with PCR showed a specificity & sensitivity of 85.7% and 85.7%, respectively for visual detection of high risk HPV detection. 32 Majority of the reported studies on colorimetric analysis for biosensing have deployed Au NPs that were pre-formed. The formation of Au NPs is highly dependent on various factors that triggers or hinders their growth.…”