Multiplexed colorimetric detection of Kaposi's sarcoma associated herpesvirus and Bartonella DNA using gold and silver nanoparticles

Abstract

Kaposi’s sarcoma (KS) is an infectious cancer occurring most commonly in human immunodeficiency virus (HIV) positive patients and in endemic regions, such as Sub-Saharan Africa, where KS is among the top four most prevalent cancers. The cause of KS is the Kaposi’s sarcoma-associated herpesvirus (KSHV, also called HHV-8), an oncogenic herpesvirus that while routinely diagnosed in developed nations, provides challenges to developing world medical providers and point-of-care detection. A major challenge in the diagnosis of KS is the existence of a number of other diseases with similar clinical presentation and histopathological features, requiring the detection of KSHV in a biopsy sample. In this work we develop an answer to this challenge by creating a multiplexed one-pot detection system for KSHV DNA and DNA from a frequently confounding disease, bacillary angiomatosis. Gold and silver nanoparticle aggregation reactions are tuned for each target and a multi-color change system is developed capable of detecting both targets down to levels between 1 nM and 2 nM. The system developed here could later be integrated with microfluidic sample processing to create a final device capable of solving the two major challenges in point-of-care KS detection.

Simultaneous Detection of Dual Nucleic Acids Using a SERS-Based Lateral Flow Assay Biosensor

“…These LOD values are at least 10,000 times more sensitive than those obtained from the nanoparticle aggregation-based colorimetric method. 28 Importantly, our SERS-based LFA strip can be used for simultaneous detection of dual DNA markers, KSHV and BA, with high sensitivity.…”

“…28 In this approach, gold and silver nanoparticles aggregations were tuned for each target DNA and a multicolor changing system was developed for the detection of both targets with sensitivity of 1 nM. This aggregation-based colorimetric detection has some merits including its speed and simplicity but further improvements in sensitivity and the capability for quantitative analysis are still needed for the early diagnosis of KS and BA.…”

Gold Nanoparticles for DNA/RNA-Based Diagnostics

“…The color readout discards the use of complex equipment that makes it suitable for POC assays. More recently, this same approach was used for the naked eye and direct (amplification-free) detection of Kaposi's sarcoma-associated herpes virus [17] and Salmonella sp. [16].…”

Multiplex DNA Biosensor for Viral Infection Diagnosis Using SERS Molecular Sentinel-on-Chip

“…Among diagnostic techniques, molecular diagnosis based on nucleic acid biomarker (DNA or RNA) detection offers many advantages such as high specificity, low limit of detection, and the ability to quantify the level of infection [1]. Various multiplex DNA/RNA detection techniques have been reported [2][3][4][5][6][7][8][9].…”