---
title: 3 Applications for Highly Discriminating (HiDi®) Enzymes
description: Explore Medix Biochemica's HiDi® enzymes for advanced PCR testing, mutation detection, genotyping, cancer diagnostics, and CRISPR-Cas SNP quality control.
---

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# [3 Applications for Highly Discriminating (HiDi®) Enzymes](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes)

 Written by [Medix Biochemica](https://articles.medixbiochemica.com/author/medix-biochemica) | Aug 17, 2023 2:34:43 PM

HiDi® (pronounced “Heidi”) stands for highly discriminating.[1](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References) A highly discriminating  enzyme is an enzyme that’s been artificially modified to work in highly selective PCR testing and mutation detection.[1](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References) They are the gold standard for cases where a researcher or healthcare provider is tasked with ‘finding a needle in a haystack’, delivering a level of certainty that other products can’t provide.[1](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References) 

What Are HiDi® Enzymes and What Sets Them Apart?

A HiDi® enzyme differentiates between primers that bind to their intended targets versus primers that are not fully binding.[1](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References) In simple terms, the enzyme discards any mismatched primers and only ‘searches’ for the primers that are fully matched to the enzyme’s target sequences.[1 ](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

Many enzymes on the market today feature low discrimination or none at all.[1 ](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)This becomes an issue when tests need to focus on specific primers, e.g. in as-PCR testing.[1 ](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

Highly discriminating enzymes, on the other hand, are exceptionally useful in these cases.[1](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

[Click here for a complete list of abbreviation definitions](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#Abbreviations)

## 3 Applications for HiDi® enzymes:

**1. Genotyping**

HiDi® enzymes can be used in genotyping to determine the alleles (genetic variants) an individual might have inherited.[1-3](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

HiDi® gives testers key advantages in genotyping:[2](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

- Up to 30% total time and labor cost reduction compared to PCR- and NGS-based genotyping
- No significant protocol changes required
- Enables confident acceleration of research

**2. Cancer mutation detection**

HiDi® products can be used in as-PCR testing to detect cancer mutations.[1,2 ](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

The traditional PCR workflow would include primer design, followed by PCR testing, followed by separation of DNA fragments (gel electrophoresis).[2,4](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References) With HiDi®, the workflow is shorter, starting with primer design and progressing straight to quantitative (real-time) PCR.[2](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References) qPCR can simultaneously amplify and detect the target DNA.[5](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

**3. CRISPR-Cas SNP quality control*  
***SNPs can provide crucial information about an individual’s genetic makeup and indicate their predisposition towards certain diseases.[2 ](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)SNPs introduced by CRISPR technology must undergo quality control filtering which can be done by HiDi very efficiently.[1](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

The traditional CRISPR workflow for SNP quality control takes up to 60 days, but the workflow using HiDi® DNA polymerase only takes just over 40 days.[2](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

This means:[2](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References) 

- 20% less time required
- 20% labor cost reduction
- 60% overall total workflow cost reduction
- 50% less plastic consumable waste used
- 65% lower cost associated with external services

## **HiDi****®** Applications in HLA Typing and Matching and in Liquid Biopsies

HiDi® enzymes are valuable in HLA typing and matching, helping to test patients and donors for bone marrow and cord blood transplant compatibility.[1,6](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

HiDi® enzymes can also be used in liquid biopsies to detect mutations in a high background of wild-type or ‘irrelevant’ biotype sequences.[1,2](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References) This is useful for tracking the progress of a disease and testing the efficacy of treatment, e.g. the effect of chemotherapy on cancer progression.[1](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

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## **HiDi**® Enzymes From [Medix Biochemica](https://www.medixbiochemica.com/)

There are 4 proprietary HiDi® formulations in [Medix Biochemica’s](https://www.medixbiochemica.com/) HiDi® product family. These include enzymes, Taq enzymes and ready-to-use master mixes:[2](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

- [HiDi® DNA polymerase](https://www.medixbiochemica.com/hidir-dna-polyme-9001s.html)
- [HiDi® Taq DNA polymerase](https://www.medixbiochemica.com/hidir-taq-dna-po-9201s.html)
- [HiDi® 2x PCR Master Mix](https://www.medixbiochemica.com/hidir-2x-pcr-mas-9101s.html)
- [HiDi® Taq 2x PCR Master Mix](https://www.medixbiochemica.com/hidir-taq-2x-pcr-4200s.html)

## **What can you expect from Medix Biochemica’s HiDi****®**** products?**[2](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

- **Excellent SNP discrimination:** Differentiate alleles reliably.
- **Superior performance: **Excellent for as-PCR and the first choice for highly selective PCRs. Outperform competitors.
- **Easy primer design: **Standard primers, changing only the 3’ nucleotide.
- **Highly sensitive: **Fewer than 10 copies of a mutation can be detected in a background of >10 000 wild-type copies.
- **No false amplification:** Aptamer-based hot-start prevents false amplification and provides a fast-start function.

## **Benchmarking data for HiDi****®**** Taq DNA polymerase**

[Benchmarking data ](https://articles.medixbiochemica.com/hubfs/HiDiTaq%20benchmarking%20data.pdf)clearly shows the benefits of Medix Biochemica’s HiDi® Taq DNA polymerase.[7 ](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

This highly selective DNA polymerase variant is specially evolved for all assays where high single nucleotide discrimination is required, e.g. as-PCR, mutation detection and genotyping.[7](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

HiDi® Taq DNA polymerase efficiently discriminates matched primers from those that have a mismatch at the 3'-terminus.[7](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References) It outperforms competitor products in both signal generation and discrimination.[7](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

[**Click here**](https://articles.medixbiochemica.com/hubfs/HiDi_2x_PCR_poster.pdf) to find out how the Medix Biochemica HiDi® 2x PCR Master Mix performed compared to competing products.[8](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References)

HiDi® enzymes are the gold standard for high discrimination as-PCR testing, liquid biopsy and HLA typing and genotyping.[2](https://articles.medixbiochemica.com/3-applications-for-highly-discriminating-hidi-enzymes?hs_amp=true#References) Medix Biochemica is proud to offer HiDi® products of superior quality.

| **Abbreviations** | **Definition** |
| --- | --- |
| as-PCR | allele-specific PCR |
| CRISPR | clustered regularly interspaced short palindromic repeats |
| CRISPR-Cas | CRISPR-associated protein |
| DNA | deoxyribonucleic acid |
| HLA | human leukocyte antigen |
| IVD | in vitro diagnostics |
| NGS | next generation sequencing |
| PCR | polymerase chain reaction |
| qPCR | quantitative (real-time) PCR |
| SNP | single-nucleotide polymorphism |
| Taq DNA polymerase | thermostable DNA polymerase |

References:

1. Expert opinion. Interview with Ramon Kranaster, Managing Director and R&D Director at myPOLS Biotec GmbH. May 16, 2023.  
2. Data on file. Medix Biochemica. HiDi® Family: True gold-standard for high discrimination allele-specific PCR. February 14, 2023.
3. Karch SB. Autopsy: molecular. In: Payne-James J, Byard RW, eds. Encyclopedia of Forensic and Legal Medicine (Second Edition). Elsevier; 2016:290-296. doi:10.1016/B978-0-12-800034-2.00041-0.
4. Polymerase chain reaction (PCR). Khan Academy. Accessed June 13, 2023. [https://www.khanacademy.org/science/ap-biology/gene-expression-and-regulation/biotechnology/a/polymerase-chain-reaction-pcr](https://www.khanacademy.org/science/ap-biology/gene-expression-and-regulation/biotechnology/a/polymerase-chain-reaction-pcr).
5. Hussain B, Yüce M, Ullah N, et al. 3 - Bioconjugated nanomaterials for monitoring food contamination. In: Grumezescu AM, ed. Nanobiosensors. Academic Press; 2017:93-127. doi:10.1016/B978-0-12-804301-1.00003-5.
6. HLA basics. BeTheMatch.org. Published March 10, 2023. Accessed June 13, 2023. [https://bethematch.org/transplant-basics/how-blood-stem-cell-transplants-work/hla-basics/](https://bethematch.org/transplant-basics/how-blood-stem-cell-transplants-work/hla-basics/).
7. Data on file. Medix Biochemica. [Benchmarking data for HiDi® Taq DNA polymerase (#9201).](https://articles.medixbiochemica.com/hubfs/HiDiTaq%20benchmarking%20data.pdf)
8. Data on file. [Medix Biochemica 33rd ECCMID. 2023.](https://articles.medixbiochemica.com/hubfs/HiDi_2x_PCR_poster.pdf) 

 

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