Abstract
The TMPRSS2:ERG gene fusion is one of a series of highly promising prostate cancer (PCa) biomarker alternatives to the controversial serum PSA. Current methods for detecting TMPRSS2:ERG are limited in terms of long processing time, high cost and the need for specialized equipment. Thus, there is an unmet need for less complex, faster, and cheaper methods to enable gene fusion detection in the clinic. We describe herein a simple, rapid and inexpensive assay which combines robust isothermal amplification technique with a novel visualization method for evaluating urinary TMPRSS2:ERG status at less than USD 5 and with minimal equipment. The assay is sensitive, and rapidly detects as low as 105 copies of TMPRSS2:ERG transcripts while maintaining high levels of specificity.
Highlights
Current prostate cancer (PCa) screening relies mainly on measuring serum prostate-specific antigen (PSA) levels which has clearly demonstrated improvements in patient survival[1,2,3]
Reverse transcription-polymerase chain reaction (RT-PCR) is a sensitive assay which is routinely used in many studies for detecting low levels of TMPRSS2:ERG transcripts in PCa urine specimens[15,16,17]
The successful detection of TMPRSS2:ERG in DuCap cells demonstrated the specificity of our assay to detect TMPRSS2:ERG transcripts from total RNA
Summary
Current prostate cancer (PCa) screening relies mainly on measuring serum prostate-specific antigen (PSA) levels which has clearly demonstrated improvements in patient survival[1,2,3]. PSA screening has been utilized widely for several decades, its value as a biomarker is controversial due to its poor sensitivity and specificity As a consequence, this has frequently led to over diagnosis and unnecessary medical expenses[1,4,5]. RT-PCR is a sensitive assay which is routinely used in many studies for detecting low levels of TMPRSS2:ERG transcripts in PCa urine specimens[15,16,17]. DNA)-mediated flocculation assays[20,21,22] which typically possess a binary threshold for flocculation output and interpretation[23,24,25] This sort of binary evaluation system would complement the binary nature of the TMPRSS2:ERG biomarker. The assay is a relatively simple, rapid, and inexpensive methodology for evaluating urinary TMPRSS2:ERG status with good sensitivity and specificity
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