| CPC C12Q 1/6851 (2013.01) [B01L 3/502715 (2013.01); B01L 3/50273 (2013.01); B01L 3/5457 (2013.01); C12Q 1/686 (2013.01); B01L 2300/021 (2013.01); B01L 2300/042 (2013.01); B01L 2300/0645 (2013.01); B01L 2300/0654 (2013.01); B01L 2300/18 (2013.01); B01L 2400/0415 (2013.01); B01L 2400/0439 (2013.01); B01L 2400/082 (2013.01)] | 20 Claims |
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1. A reverse transcriptase quantitative polymerase chain reaction based analyzing system comprising:
a sampling tube comprising:
a tube portion;
a cap coupled to the tube portion via a ratchet locking mechanism, wherein the cap comprises a lens; and
a radio frequency identification (RFID) tag disposed between the cap and the tube portion; and
a microfluidic processing unit coupled to the sampling tube, wherein the microfluidic processing unit comprises:
a piezo electric type element, electrostatic ribonucleic acid (RNA) extraction and concentration unit coupled to the sampling tube via a plurality of sample flow control devices;
an eluent storage unit coupled to the piezo electric type element, electrostatic ribonucleic acid extraction and concentration (REC) unit via a plurality of eluent flow control devices;
an eluate dosing chamber coupled to the REC unit via a plurality of eluate flow control devices;
an analysis settling chamber coupled to the eluate dosing chamber;
an assay rehydration unit coupled to the analysis settling chamber via a plurality of assay control devices;
an assay analysis unit coupled to the analysis settling chamber; and
a no-target control analysis unit coupled to the assay rehydration unit for the purposes of applying a negative control to the biological assay and simultaneously calibrating the optical metrology system by the provision of passive dyes that will be present in effectively the same quantities in both the no-target control and the biological sample, wherein the use of lyophilization in a factory to leave the required reagents in the allocated chambers of the microfluidic processing unit wherein the final stage of preparation prior to calibration involves displacing air in channels and chambers by argon in order to dispel oxygen that might otherwise facilitate gradual deterioration in the regents by oxidation.
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15. A method for analyzing a biological sample, using a reverse transcriptase quantitative polymerase chain reaction analyzing system, the method comprising:
receiving a sample within a tube portion of a sampling tube;
coupling a cap of the sampling tube to the tube portion via a ratchet locking mechanism;
depositing a microfluidic laboratory comprising the sampling tube in an analytic cell (AC), wherein AC is an analytical process environment, wherein the AC may be just a single unit or may be one of a number assembled in a row; wherein a row of ACs, known as a RandOm-Access Analytical (RoaaaR) Array configured as one of a singular unit or one of many housed within an outer chassis, wherein the deposition is by way of either manual insertion or robotic assignment of individual sampling tubes and microfluidic laboratories into vacant ACs; wherein asynchronous, simultaneous processing of sampling tubes integrated with microfluidic laboratories whereby each sample test commences as it is inserted into an AC; wherein a control system to maintain asynchronism between assays in progress wherein optical metrology system is kept in constant use without delaying any given assay; wherein a signal detection mechanism using modulation of the excitation light correlated with both the spectrum and the intensity of individual wavelengths in the emission spectra to reduce electrical, electronic, optical and biological noise; wherein use of same modulation of the excitation light as a means of multiplexing by way of time division multiplexing (TDM) emissions from two or more ACs to economically use the same optical metrology system;
checking, by one or more detection sensors, the locking of the cap to the tube portion;
mixing a lysis buffer with the sample within the sampling tube to generate a lysated sample within the sampling tube via a mixing unit;
circulating the lysated sample from the sampling tube to a piezo electric type element, electrostatic ribonucleic acid extraction and concentration unit via a plurality of sample flow control devices;
separating ribonucleic acid strands from the lysated sample within the piezo electric type element, electrostatic ribonucleic acid extraction and concentration unit;
circulating an eluent between an eluent storage unit and the piezo electric type element, electrostatic ribonucleic acid extraction and unit;
extracting the separated ribonucleic acid strands, using the eluent from the piezo electric type element, electrostatic ribonucleic acid extraction unit and transferring an eluate comprising the extracted ribonucleic acid strands to an analysis settling chamber via an eluate dosing chamber and a plurality of eluate flow control devices;
mixing a portion of master mixture from an assay rehydration unit with the eluate within the analysis settling chamber;
transferring a first portion of the master mixture and the eluate to an assay analysis unit to analyze mixture of the portion of the master mixture and the eluate to diagnose a biological condition associated with the sample;
transferring a second portion of the master mixture from the assay rehydration unit to a no-target control analysis unit to perform a no-target control analysis of the second portion of the master mixture;
performing at least one of uniform (isothermal) or varying temperature control of the reverse transcriptase, quantitative polymerase chain reaction (RT-qPCR) assay or any other assay chemistry benefitting from the high accuracy, contaminant-free, optical detection methods;
providing asynchronous processing of the collection of simultaneous assays in a system to minimize the test extraction periods and share the more costly resources such as the optical subsystem between assays without slowing individual assays; and
equipping individual microfluidic laboratories with mirrors underneath their reaction chambers to maximize the available emitted fluorescence, approximately doubling the light at each stage and for qPCR or RT-qPCR, reducing the cycle count by one.
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