User:TemPCR

The History of Molecular Based RT-PCR and Molecular Testing
For the past 25 years RT-PCR has been used as a molecular based testing platform to accurately identify a single pathogen. RT-PCR has served the medical community as a valuable tool for confirmation testing but has had several technology barriers that have prevented RT-PCR from becoming the clinical diagnostic tool of choice. The greatest limitation of RT-PCR testing is the inability to test for multiple pathogens in a single result. The specific testing amplification requirements for each pathogen using RT-PCR have made the process to cost prohibitive and too labor intensive to use as a stand alone testing method for clinical diagnosis. Some RT-PCR product companies have attempted to ease this problem by providing RT-PCR machines that simplify this process. Although these methods have reduced the time needed to run single a pathogen RT-PCR test, these methods have not solved the true need for multiple pathogen identification in single a molecular test.

The Problem of Molecular Multiplexing Using RT-PCR
The following is a list of common problems associated with multiplex PCR  assay development:

Incompatible Loci. Each target in a multiplex reaction demands its own optimal condition. Therefore, increasing the number of targets in a multiplex reaction can become difficult and is often impossible to execute.

Lack of Specificity. Multiple sets of high-concentration primers in a system can generate primer dimers and non-specific, background amplification.

Lack of Sensitivity. Crowding of primers reduces amplification efficiency and wastes resources by occupying enzyme and substrates.

Uneven Amplification. Differences in amplification efficiency of each target may lead to large discrepancies in amplicon yields. In a multiplex system, some loci may amplify well, while others may amplify poorly or not at all. Uneven amplification also makes it impossible to accurately perform end-point quantitative analysis.

Lot-to-lot Variation. Due to the fact that large amounts of primers are consumed in each reaction and manufacturers can generate only a limited amount of assays per lot, quality control and assurance can be difficult.

The Multiplex Innovation of TemPCR
Tem-PCR is a proprietary true molecular multiplex PCR technology that stands for Target Enriched Multiplexing. This award winning breakthrough technology overcomes the limitations of RT-PCR testing by its unique ability to identify multiple genetic targets in a single molecular test. This proprietary testing process allows for specific and sensitive amplification of multiple targets in a single tube. This unique ability of Tem-PCR to test for multiple pathogens both viral and bacterial allows for not only more accurate identification of pathogens but also Molecular Differential Diagnosis (MDD).

Like modern CT technology that can scan the human body for structural abnormalities, Tem-PCR technology can scan a DNA sample and identify the particular pathogen responsible for an infection, or detect genetic changes within that pathogen which contribute to its drug resistance. Tem-PCR is a disruptive technology, delivering incredible benefits and advances to modern medical practice.

Progressive healthcare professionals and providers recognize that personalized medicine requires personalized diagnosis. This methodology treats targeted causative agents associated with a particular patient, not the symptoms associated with a group of patients. Until Tem-PCR physicians could only prescribe treatment based on statistics, i.e., “if I give this drug, I will have 75% chance that the patient will respond favorably.” This means there is 25% of the risk that the patient may not respond to delivered treatment. Now physicians have a more accurate diagnostic tool with the ability Tem-PCR provides to use Molecular Differential Diagnosis or MDD. Using the process of MDD; Tem-PCR now makes it possible to molecularly differentiate between vial and bacterial pathogens, CA and HA infections, co-infections, pathogenic subtypes, and their associated drug resistance in a single molecular test.

Why is TemPCR so Accurate
Most of the problems associated with standard multiplex PCR are due to the presence of multiple sets of high concentration labeled primers. Yet, these high concentrations are only required in the last cycles of a PCR reaction. With Tem-PCR, we use only enough gene-specific primers to "enrich" the targets and introduce the Super Primer tag into the products. Then, amplification is carried out with only one pair of primers.

Incompatibility among co-amplified targets is another major roadblock to standard multiplex assay development. Usually, one pair of primers define each target; therefore, one optimal condition must be identified that can allow for the co-amplification of all targets. With Tem-PCR, a target is amplified by nested primers producing four potential products for exponential amplification, thereby creating multiple optimal conditions. This feature of Tem-PCR increases compatibility among the co-amplified loci and creates fewer restrictions for multiplex assay development.

TemPCR Allows for Molecular Differential Diagnosis (MDD)
The personalized diagnostic approach that MDD provides healthcare professionals allows physicians to use to power of molecular diagnostics in an entirely new way to improve patient care. The capacity to quickly and definitively differentiate between pathogens and drug resistance not only provides the ability for faster more appropriate treatment of patients, but also prevents unnecessary patient exposure to ineffective drug compounds. The information provided to physicians through MDD truly assist the physician in improving patient outcomes and reducing unnecessary length of stay. The power of MDD makes Tem-PCR truly the first molecular diagnostic test that assist physicians with the practice of personalized medicine.