Answer: In case of assays with RNA as the nucleic acid, reverse transcription PCR (RT PCR) is performed. The Truenat pouch contains Microtube with freeze dried RT PCR reagents. Six (6) μL of the purified RNA is then dispensed using the provided micropipette and tip into the microtube containing freeze dried RT PCR reagents, including reverse transcriptase (RT) and allowed to stand for 30-60 seconds to get a clear solution. Six (6) μL of this clear solution is then pipetted out using the same pipette and tip and dispensed into the reaction well of the Truenat disease specific chip and the test is inserted in the Truelab Real Time micro PCR Analyzer where the RNA is first converted into complementary DNA (cDNA) by the RT enzyme and further thermal cycling takes place.
Answer : The Truelab Real Time quantitative micro PCR System automates sample preparation and nucleic acid amplification, and detection of the target sequence in simple and complex samples using real time polymerase chain reaction (PCR). The system is developed with the aim to bring Real Time PCR testing to a point-of-care level to enable early diagnosis and better patient care.
The Truelab Real Time quantitative micro PCR system comprises of three components:
1. Truelab Real Time Micro PCR workstation:
A. Trueprep AUTO Universal Cartridge Based Sample Prep Device:The PCR process necessitates the extraction and purification of nucleic acids from clinical specimens to free it from potential PCR inhibitors. The Trueprep AUTO Universal Cartridge Based Sample Prep Device together with Trueprep™ AUTO Universal cartridge Based Sample Prep Kit provides an easy method of nucleic acid extraction and purification.
B. Truelab Uno Dx/Duo/Quattro Real Time Quantitative micro PCR Analyzer: The Truelab Uno Dx/Duo/Quattro Real Time micro PCR analyzer performs automatic amplification of the pathogenic nucleic acids extracted from the sample and also displays result of the final analysis of the PCR/ RT- PCR run.
2. Trueprep AUTO Universal Cartridge Based Sample Prep Kit: Trueprep AUTO Universal Cartridge Based Sample Prep Kit contains cartridges and reagents for purification of nucleic acids from biological specimen. The entire process of extraction and purification of nucleic acids is completely automatic and takes about 20 minutes.
3. Truenat Chip Based Real Time PCR test: Truenat is a disposable, room temperature stable, chip with dried MgCl2 in reaction well and freeze dried PCR reagents in microtube for performing Real Time PCR test and runs on the Truelab Real Time micro PCR Analyzer.
Answer : Trueprep AUTO Universal Cartridge Based Sample Prep Kit protocol uses a proprietary matrix enclosed in a cartridge to purify nucleic acids from clinical samples. The cartridge also contains pre-loaded Internal Positive Control (IPC).The IPC is a full process control that undergoes all the processes the specimen undergoes, from extraction to amplification, thereby validating the analysis from sample to result.
Specimen pre-treated with lysis reagent is added to the sample chamber of the cartridge which is then placed in the cartridge holder of the device. Sample processing is initiated upon pressing the start button on the device, through an Automatic pre-programmed process wherein Nucleic Acids released by chemical and thermal lysis of cells bind to the proprietary matrix in the matrix chamber. In subsequent steps, the captured nucleic acids are washed with buffers to remove the PCR inhibitors and finally eluted from the matrix using the elution buffer. At the end the cartridge is Automatically ejected and the elute, containing purified nucleic acids is then collected from the elute chamber for further analysis.
Answer : Trueprep AUTO Universal cartridge based sample prep Device along with Trueprep AUTO Universal Cartridge based sample prep kits can be used to extract nucleic acids from Sputum, Whole blood, Serum, Plasma, Pus, Peritoneal Fluid, Pleural Fluid, Cerebrospinal fluid (CSF), BAL, Lymph-node Aspirate, Abscess, urine, stool, Animal tissues, saliva & different swab specimens.
Answer : Trueprep AUTO Universal Cartridge based Sample Prep Device extracts nucleic acids that can be analysed on any kind of analyzer available in the market. It is also sold as standalone Device.
Answer : There are 3 Variants available in Truelab Real Time Micro PCR analyzer. Namely,
A. Truelab Uno Dx Real Time Quantitative micro PCR Analyzer
B. Truelab Duo Real Time Quantitative micro PCR Analyzer
C. Truelab Quattro Real Time Quantitative micro PCR Analyzer
Answer: Truelab Uno Dx, Duo and Quattro can perform 12, 24 and 48 tests in 8 hours, respectively.
Answer : Yes.
Answer : Ct (Threshold cycle) reflects the the cycle number at which the fluorescence signal generated within the reaction crosses the threshold. It is inversely correlated to the logarithm of the initial copy number. The Ct assigned to a particular sample thus reflects the point during the reaction at which a sufficient number of amplicons have accumulated. It is also sometimes called the crossing over (Cp) point.
Answer : No, every lot of the Truenat chip comes with the in-built Standard curve values. So when you run the Truenat chip you do not require to run a standard curve every time.