Naked eye detection of the Mycobacterium tuberculosis complex by recombinase polymerase amplification—SYBR green I assays

Abstract

A rapid, sensitive, naked eye RPA assay can be integrated into point-of-care diagnosis for Mtb detection, especially in remote areas where laboratory instrument resources are limited.

Rapid detection of extended spectrum β-lactamase producing Escherichia coli isolated from fresh pork meat and pig cecum samples using multiplex recombinase polymerase amplification and lateral flow strip analysis

“…The double-stranded DNA template is dissociated by a recombinase and single-stranded binding protein, and then, Sau DNA polymerase generates new complementary products within 5-20 min [18]. The results of alternative post-RPA detection methods using SYBR Green or a lateral flow strip assay (LFA) are easily observed by the naked eye, and can be applied for point-of-care testing and field studies [19][20][21]. Hence, this study attempted to develop multiplex RPA combined with LFA to detect the three most common ESBL genes (bla CTX-M , bla OXA , and bla SHV ).…”

Development of a reverse transcription recombinase polymerase amplification assay for rapid and direct visual detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)

“…The end product of the RT-RPA can also be detected conveniently using a LF strip. On balance, the end point detection using SYBR Green or LF strip used less instrumentation than real-time monitoring, thus reducing the overall cost of the test and suited to resource-limited settings [7]. Moreover, the test results either appear as visible bands or show a colour change that can be easily seen and interpreted with the naked eye.…”

Rapid detection of multidrug-resistant tuberculosis based on allele-specific recombinase polymerase amplification and colorimetric detection

“…A higher sensitivity of 0.05 ng was reported in an RPA coupled with SYBR-based detection of M . tuberculosis using an IS1081-specific primer in our previous study [ 16 ]. The lower sensitivity observed in this study can probably be attributed to the nature of the targeted gene detected.…”

“…RPA has been employed in many studies to detect the M . tuberculosis genome [ 13 , 16 , 34 , 35 ]. However, to the best of our knowledge, only two publications demonstrate MDR-TB detection by RPA integrated by lab-on-a-disc and a fluorescent detector [ 14 , 15 ].…”