Introduction

In molecular biology research and diagnostic applications, pathogens are becoming increasingly diverse and complex. The challenge of rapidly and accurately identifying pathogen types has intensified, and residual background microbial nucleic acids have become one of the key factors affecting the accuracy and reliability of test results.

Industry Pain Point: “Achilles’ Heel” of High Sensitivity — False Positives

Polymerase Chain Reaction (PCR)-based molecular diagnostic technologies are recognized for their high sensitivity, specificity, and efficiency. They have been recommended by multiple authoritative international and domestic guidelines, including the latest EORTC/MSGERC guidelines, as essential tools for early and rapid detection of infectious diseases — such as COVID-19, invasive pulmonary fungal disease (IPFD), and various respiratory and intestinal infections.

However, high sensitivity is a double-edged sword. Contaminating background nucleic acids can obscure low-abundance target nucleic acids or be co-amplified with the target, leading to reduced detection sensitivity or false positive results — causing confusion in clinical diagnosis and treatment. Contamination can arise during product manufacturing, storage, or use, and even from operators themselves.

 

Tracing the Source: The Key to Zero Nucleic Acid Residue Lies in Enzyme Raw Materials

For bacterial and fungal detection, false positives primarily stem from one critical raw material — enzymes. The commonly used expression systems for industrial enzyme production (such as E. coli, yeast, or Aspergillus) are themselves the very pathogens targeted by molecular detection assays. Without strict control, enzymes carrying residual nucleic acids used in diagnostic reagent production can directly cause batch-specific false positives (either through nucleic acid contamination or nonspecific amplification), severely impacting clinicians’ diagnostic accuracy and treatment decisions. Therefore, to enhance the performance of in vitro diagnostic reagents, selecting “ultra-clean” enzyme raw materials free from nucleic acid residues is essential.

Facing an industry where enzyme quality varies widely and nucleic acid residue control standards are inconsistent, Hzymes has refined and upgraded its production processes, establishing a systematic nucleic acid removal pathway and launching “ultra-clean” enzymes and MIX formulations that eliminate background contamination.

1. Strict Control of Internal and External Contamination

System: Fully compliant with GMP quality management, ensuring comprehensive monitoring of personnel, equipment, materials, methods, and environment.
Facilities: Rationally designed production and testing spaces with strict physical segregation to prevent cross-contamination.
Materials: All production materials and consumables undergo the most stringent selection and quality control, cutting off contamination from the source.

2. Efficient Nucleic Acid Removal at the Front-End

Multi-method purification: In the initial purification phase, a combination of heat treatment, precipitation, enzymatic digestion, and proprietary purification partners is employed to efficiently remove the majority of free nucleic acid contaminants.

3. Deep Purification via Multi-Mode Chromatography

Orthogonal chromatography strategy: A strategically optimized combination of heparin, anion-exchange, gel filtration, hydrophobic interaction, and composite chromatographic methods.
Extreme separation performance: Through multi-step orthogonal chromatography, nucleic acid residues are thoroughly removed based on charge, size, and hydrophobicity differences between nucleic acids and target proteins.

4. Complete Terminal Nucleic Acid Inactivation

Targeted enzymatic digestion and inactivation: Highly efficient nucleases that do not affect the target protein are carefully selected, and digestion conditions are finely optimized. Following purification, the enzyme-treated products undergo rigorous inactivation validation to ensure complete removal of the nucleases themselves, guaranteeing absolute cleanliness and safety of the final product.

The Ultra-Clean Choice — Hzymes MIX Evaluation Data

 

1. Using HMD3818 to detect E. coli and Aspergillus, all 48 wells tested negative.

1. Using HMD3813 to detect E. coli, Acinetobacter baumannii, and Pseudomonas aeruginosa, all 48 wells tested negative.

 

1. Using HMD3808 to detect E. coli genomic DNA — in negative samples, Hzymes reagent: 0/50 positives; competitor reagent: 3/10 positives (blue = positive; red = negative).

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