Significant Expansion of Real-Time PCR Multiplexing with Traditional Chemistries using Amplitude Modulation
| العنوان: | Significant Expansion of Real-Time PCR Multiplexing with Traditional Chemistries using Amplitude Modulation |
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| المؤلفون: | Dominic Yurk, Karen L. Menge, Claudia Shin, Scott E. Fraser, Aditya Rajagopal, Gregory J. Tsongalis, Thomas A. Tombrello, Lucien Jacky |
| المصدر: | Scientific Reports, Vol 9, Iss 1, Pp 1-8 (2019) Scientific Reports |
| بيانات النشر: | Springer Science and Business Media LLC, 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | 0301 basic medicine, Multidisciplinary, Nucleic acid quantitation, Channel (digital image), lcsh:R, Reproducibility of Results, lcsh:Medicine, Single sample, Real-Time Polymerase Chain Reaction, Multiplexing, Article, Amplitude modulation, 03 medical and health sciences, 030104 developmental biology, 0302 clinical medicine, Real-time polymerase chain reaction, Nucleic Acids, TaqMan, lcsh:Q, lcsh:Science, Biological system, 030217 neurology & neurosurgery |
| الوصف: | The real time polymerase chain reaction (rtPCR) is an essential method for detecting nucleic acids that has a wide range of clinical and research applications. Current multiplexed rtPCR is capable of detecting four to six nucleic acid targets in a single sample. However, advances in clinical medicine are driving the need to measure many more targets at once. We demonstrate a novel method which significantly increases the multiplexing capability of any existing rtPCR instrument without new hardware, software, or chemistry. The technique works by varying the relative TaqMan probe concentrations amongst targets that are measured in a single fluorometric channel. Our fluorescent amplitude modulation method generates a unique rtPCR signature for every combination of targets present in a reaction. We demonstrate this technique by measuring nine different targets across three color channels with TaqMan reporting probes, yielding a detection accuracy of 98.9% across all combinations of targets. In principle this method could be extended to measure 6 or more targets per color channel across any number of color channels without loss in specificity. |
| وصف الملف: | application/pdf |
| تدمد: | 2045-2322 |
| DOI: | 10.1038/s41598-018-37732-y |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3ee6ced1756cbfb277cfba9635c08c77 https://doi.org/10.1038/s41598-018-37732-y |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....3ee6ced1756cbfb277cfba9635c08c77 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 20452322 |
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| DOI: | 10.1038/s41598-018-37732-y |