COVID-19 ONE-STEP RT-PCR KIT

What is RT- PCR?

Reverse transcription polymerase chain reaction (RT-PCR) is a standard PCR variant that consists of amplified forms of particular mRNA, attained from minute samples. It eradicates the requirements for the burdensome mRNA purification procedure needed in the traditional cloning practices. In RT-PCR, in addition to the standard PCR reagents, reverse transcriptase and an RNA sample are used. The reaction mixture is heated to a temperature of 37 ˚C, which permits cDNA production from the RNA sample through the mechanism of reverse transcription. This cDNA hardens to one of the primers leading to first-strand synthesis. Standard PCR continues, producing dsDNA.

The primers used in the procedure must counterpart the project. Oligo (dT) primers are sufficient in case amplification of the entire mRNA of a cell is required because they get attached to the poly(A) tails. On the contrary, if a particular mRNA needs to be amplified, a primer specific to the coding region can be used.

RT-PCR is a general virology diagnostic procedure, often combined with quantitative real-time PCR (qPCR), which is extensively used for quantification of RNA transcript levels in cells and tissues. The combination of real-time PCR (qPCR) and reverse transcription PCR is referred as quantitative RT-PCR or qRT-PCR

 

What is Multiplex PCR?

Multiplex polymerase chain reaction refers to the use of polymerase chain reaction to amplify several different DNA sequences simultaneously. This process amplifies DNA in samples using multiple primers and a temperature-mediated DNA polymerase in a thermal cycler.

Meril COVID-19 one Step RT-PCR Kit

Due to the novel coronavirus (SARS-CoV-2) outbreak, Meril Diagnostics has worked extensively to develop a new one step multiplex real-time RT-PCR kit to allow laboratories to diagnose COVID-19 caused by SARS-CoV-2 infection with fast and easy to use assays. Meril Covid-19 one step Rt PCR kit is a fast, highly sensitive multiplex diagnostic kit that contains both the assays and controls needed for the real-time PCR detection of RNA from the SARS-CoV-2 virus.

It is a One tube Multiplex PCR reaction for identification and detection of 2019-ncov.  Its Primer and Probe mix based on dual-target gene design the two confirmatory gene ORF 1ab gene and nucleoprotein N gene for the detection of Sars COV 2 in human sample, with an analytical Analytical Accuracy of < 5 RNA copies/ reaction it makes Meril COVID-19 one Step RT_PCR Kit a highly sensitive and specific kit for the detection of SARS COV 2 RNA.

The kit is highly compatible with Open ended RT PCR instruments with FAM, HEX/VIC, RED/ROX channels. The easy protocol, minimum assay time and high specificity and sensitive makes it an ideal choice for your laboratory for the detection of COVID 19 suspected patients.

Meril COVID-19 RT-PCR Kit compatible with popular Applied Biosystems (ABI) 7500, BioRad CFX 96, Shanghai Hongshi SLAN-96P, QIAGEN Rotor Gene Q  and other PCR instruments with FAM, HEX/VIC, RED/ROX channels.

Principle: 

The primer and probe mix for this kit accepts the dual-target gene design, which aims the particularly conserved sequence encoding the ORF 1ab gene and the nucleoprotein N gene. The template’s amplification can be quantitatively monitored by the increasing fluorescence signal detected by a real time PCR instrument, as the PCR reaction mix is readily provided.

The PCR detection system comprises of an endogenous internal control primer and probe mix. The result of internal control provides precise sampling and extraction process, to avoid false negative results.

Intended Use:

This kit is designed to detect COVID-19 disease using real time PCR. The results can be used to assist diagnosis of patients with COVID-19 infection, and provide molecular diagnostic basis for infected patients.

This RT-PCR kit can be utilized by clinical and public health laboratories to rapidly assess up to 94 patient specimens with a single kit, within a span of 3 hours. The kit is approved for use with RNA taken out from nasopharyngeal swabs, nasopharyngeal aspirate (nasal aspirate), and bronchoalveolar lavage (BAL) from patients at risk of getting exposed to the SARS-CoV-2 virus or having signs and symptoms of COVID-19.

Results are for recognizing SARS-CoV-2 RNA. During the acute phase of infection, the SARS-CoV-2 RNA is usually evident in any of the above specimens. Positive results are suggestive of the SARS-CoV-2 RNA presence; albeit, clinical link with patient history and other diagnostic information is essential to decide the stage of patient infection. Positive results do not exclude bacterial infection or co-infection with other viruses. The identified agent may not be the certain cause of disease.
Also, negative results do not disqualify SARS-CoV-2 infection and hence, should not be used as the only basis for deciding patient management. Negative results have to be pooled with clinical observations, patient history, and epidemiological information.

Evaluation with the Meril COVID-19 One Step RT PCR Kit is meant for use by qualified and trained clinical laboratory personnel, specially instructed and trained in the real-time PCR procedures and in vitro diagnostic procedures.

The test results of this kit are meant for clinical reference only and should not be used as the sole reference for clinical diagnosis. Conducting an inclusive analysis is suggested by combining the test results with patients’ symptoms and other laboratory tests.

 

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