PCR Technology: Evolution and Modern Applications

At a Glance

Polymerase chain reaction (PCR) has become one of the most valuable techniques currently used in bioscience, diagnostics and forensic science [3]. This article traces the development of PCR from its invention to current variants and compares their applications in diagnostics and research. The table below summarizes the main PCR technology generations and their primary uses.

Historical Development of PCR

Early Foundations

The genetic information that dictates the structure and function of all life forms is encoded in DNA. In 1953, Watson and Crick first presented the double helical structure of a DNA molecule [4]. Their findings unearthed the desire to elucidate the exact composition and sequence of DNA molecules. In 1959, Kornberg reproduced the replication process of DNA in a test tube [17]. The discovery of Taq polymerase in 1969 provided a heat-stable enzyme essential for the PCR process [17].

Invention and Proof of Concept

PCR began in the late 1960s and early 1970s for the study of gene in vitro isolation technology [17]. In 1984, PCR was proven to be feasible [17]. The original method used a DNA polymerase from E. coli that was inactivated during the denaturation step, requiring fresh enzyme addition each cycle. The introduction of Thermus aquaticus (Taq) polymerase, which remains stable at high temperatures, automated the process and improved specificity and yield.

Commercialization and Widespread Adoption

Improvements such as the use of fluorescent dyes and the implementation of PCR for amplification led to sequencing a few hundred base pairs in days, to automation where sequencing of thousands of base pairs in hours became possible [10]. Currently, there are two main areas of PCR utilization in bioscience: high-throughput PCR systems and microfluidics-based PCR devices for point-of-care (POC) applications [3].

Core Principles of PCR

Thermal Cycling

PCR relies on repeated temperature changes to amplify specific DNA sequences. Each cycle consists of three steps: denaturation (94-98°C) to separate double-stranded DNA, annealing (50-65°C) for primer binding, and extension (72°C) for DNA synthesis by Taq polymerase. The number of target DNA molecules doubles approximately with each cycle, yielding exponential amplification.

Primer Design

Primers are short oligonucleotides that flank the target sequence. Successful amplification depends on primer specificity, melting temperature compatibility, and absence of secondary structures. Universal PCR primers have been developed for applications such as metabarcoding environmental DNA from fishes, targeting a hypervariable region of the 12S rRNA gene [8].

Enzyme Selection

Taq polymerase remains the standard enzyme for routine PCR. High-fidelity polymerases with proofreading activity are used when sequence accuracy is critical, such as in cloning or mutation detection. The choice of enzyme affects amplification efficiency, error rate, and product length.

Real-Time Quantitative PCR (qPCR)

Principles and Chemistries

Real-time PCR, also known as quantitative PCR (qPCR), monitors amplification in real time using fluorescent dyes or probes [20]. Two main chemistries exist: DNA-binding dyes such as SYBR Green that fluoresce when bound to double-stranded DNA, and sequence-specific probes such as TaqMan that generate fluorescence only when the target is amplified.

Quantification Methods

Absolute quantification requires a standard curve generated from known concentrations of target DNA. Relative quantification compares target gene expression to a reference gene. The cycle threshold (Ct) value, the cycle at which fluorescence exceeds background, is inversely proportional to the initial target quantity.

Applications in Diagnostics

Real-time reverse transcription PCR is the gold standard for SARS-CoV-2 diagnoses [3]. It can also be used for POC applications, being a key component of the sample-to-answer system [3]. In chronic myeloid leukemia monitoring, the 1copy BCR::ABL1 qPCR assay demonstrated reliable analytical performance and high concordance with established quantitative assays at clinically relevant molecular response thresholds [12].

Quality Control Considerations

Precision, linearity, and detection capability should be assessed according to Clinical and Laboratory Standards Institute guidelines [12]. Coefficients of variation at different concentration levels provide information about assay reproducibility. Standard curves should cover the expected dynamic range of clinical samples.

Digital PCR (dPCR)

Principle of Partitioning

Digital PCR partitions the sample into thousands to millions of individual reactions, each containing zero or one target molecule. After amplification, the proportion of positive partitions is used to calculate the absolute target concentration without requiring a standard curve [22].

Advantages Over qPCR

Digital PCR provides absolute quantification and is less affected by PCR inhibition. It offers higher precision for detecting rare mutations and copy number variations. The technology has been applied in precise molecular diagnosis, though opportunities and challenges remain [23].

Limitations

Digital PCR has a limited dynamic range compared to qPCR, typically spanning 4-5 logs versus 6-8 logs for qPCR. The cost per sample is higher, and the technology requires specialized instrumentation. Partition volume and number affect quantification accuracy.

Applications in Diagnostics

Infectious Disease Detection

Nucleic acid amplification technology (NAT) testing for blood donor screening was driven by the AIDS and hepatitis C virus epidemics, with thousands of recipients of infected blood products and components [7]. German transfusion centers were the first to start in-house NAT testing of their donations in pools of up to 96 samples for HCV, HBV, and HIV-1 [7]. Currently, more than 60 million donations per year are NAT tested worldwide, and the remaining residual risk of virus transmission by blood components and products has been reduced to almost zero [7].

When severe acute respiratory syndrome coronavirus and West Nile virus emerged, NAT enabled manufacturers and transfusion centers to instantly introduce sensitive and specific screening tests [7]. Subsequent automation including sample preparation has significantly reduced the costs and complexity of the procedure and made it affordable to middle-income countries as well [7].

Genetic Disorder Screening

Advances in molecular technologies have improved diagnostic sensitivity and specificity for hemoglobinopathy carrier detection [15]. Conventional hematological approaches, particularly complete blood count and hemoglobin typing, remain essential and cost-effective first-line tools [15]. Key challenges include unequal access to advanced diagnostics and the lack of standardized protocols across regions [15].

Cancer Monitoring

Pathogenic germline variants associated with different HER2 expression among breast cancer patients have been evaluated using next-generation sequencing-based multigene panels [13]. The 71 identified pathogenic variants were categorized into five distinct pathway clusters: BRCA/FANC, DDR, HRR, FANC, and Other [13]. The distribution of pathogenic variants enriched within these clusters differed significantly between breast cancer patients and unaffected individuals [13].

Ancient Disease Diagnosis

Diagnosis of ancient infectious diseases has seen remarkable advances, contributing to understanding historical health challenges and shaping medical research [9]. Pioneering diagnostic methods included paleopathological analysis of remains of bones and tissue lesions [9]. Later, microbiology methods were adapted to ancient materials, including molecular and serological techniques such as PCR, serology, and immunodetection, which were employed to detect DNA or antibodies from ancient pathogens under the name of the paleomicrobiology approach [9].

Applications in Research

Environmental DNA Metabarcoding

Universal PCR primers for metabarcoding environmental DNA from fishes were designed using aligned whole mitochondrial genome sequences from 880 species, supplemented by partial mitogenome sequences from 160 elasmobranchs [8]. The primers target a hypervariable region of the 12S rRNA gene (163-185 bp), which contains sufficient information to identify fishes to taxonomic family, genus, and species except for some closely related congeners [8].

Out of 180 marine fish species contained in four tanks with reference sequences in a custom database, 168 species (93.3%) were detected, distributed across 59 families and 123 genera [8]. These fishes are taxonomically diverse, ranging from sharks and rays to higher teleosts, and are greatly varied in their ecology, including both pelagic and benthic species living in shallow coastal to deep waters [8].

Plant Genetics and Breeding

Comparative plastid genome analysis of four different variants of Gastrodia elata, an important medicinal plant, revealed that the 11 plastid genomes range in size from 35,275 to 35,328 base pairs and all encode 20 protein-coding genes, 3 rRNA genes, and 5 tRNA genes [11]. Based on comparative analysis of the plastid genomes, two insertion-deletion loci were identified and corresponding InDel markers were developed, designated as GEpt11 and GEpt13 [11]. These markers were validated via PCR amplification experiments on 22 samples [11].

Wildlife Disease Surveillance

Chronic wasting disease (CWD) is a highly transmissible spongiform encephalopathy impacting cervids. Variations in the prion protein gene (PRNP), encoding the prion protein (PrP), influence disease progression and susceptibility [16]. Sequencing PRNP in white-tailed deer has revealed non-synonymous polymorphisms impacting disease [16]. Previous PRNP amplification protocols misclassified homozygous and heterozygous deer due to allelic dropout by a widely used forward primer [16]. Resequencing 586 deer previously classified as homozygous using a redesigned primer found 128 heterozygotes [16].

Workflow and Practical Considerations

Sample Preparation

DNA extraction quality directly affects PCR success. Inhibitors such as heme, humic acids, and ethanol can reduce amplification efficiency. For blood samples, automated extraction platforms have reduced costs and complexity [7]. For environmental samples, specialized extraction protocols may be required to remove inhibitors.

Reaction Setup

PCR reactions include template DNA, primers, nucleotides, buffer, and DNA polymerase. Master mixes containing all components except template reduce pipetting errors and contamination risk. Positive and negative controls should be included in every run.

Thermal Cycling Parameters

Denaturation temperature and time depend on template GC content and polymerase stability. Annealing temperature is typically 3-5°C below primer melting temperature. Extension time depends on amplicon length, with most polymerases synthesizing approximately 1 kb per minute.

Detection and Analysis

Conventional PCR products are analyzed by gel electrophoresis. qPCR data are analyzed using software that calculates Ct values and generates standard curves. Digital PCR data are analyzed using Poisson statistics to calculate target concentration.

Common Failure Patterns and Troubleshooting

No Amplification

Possible causes include degraded template, inhibitors in the sample, incorrect primer annealing temperature, or inactive polymerase. Check template integrity by gel electrophoresis or spectrophotometry. Include a positive control to verify reagent function.

Nonspecific Products

Primer-dimer artifacts or amplification of unintended targets may occur. Increase annealing temperature, redesign primers, or use hot-start polymerases. Gradient PCR can identify optimal annealing temperature.

Poor Reproducibility

Inconsistent pipetting, template degradation, or thermal cycler temperature variation can cause variability. Use master mixes, calibrate pipettes regularly, and verify thermal cycler performance. Include replicate reactions to assess precision.

Inhibition

Samples containing heme, heparin, or other inhibitors may show reduced amplification. Dilute template, use inhibitor-resistant polymerases, or purify DNA using alternative methods. Include an internal positive control to detect inhibition.

Biosafety and Laboratory Practices

Laboratory Biosafety Manual

The World Health Organization Laboratory Biosafety Manual provides guidance for safe handling of infectious materials [1]. PCR laboratories should follow appropriate biosafety levels based on the organisms being tested. Work with known pathogens requires containment facilities and practices.

Contamination Prevention

PCR is susceptible to contamination from previously amplified products. Separate pre- and post-amplification areas, use dedicated pipettes with aerosol-resistant tips, and include no-template controls. UV irradiation and bleach treatment can decontaminate surfaces and reagents.

Waste Disposal

Amplified products and contaminated materials should be disposed of according to institutional biosafety guidelines. Ethidium bromide-containing gels require special handling and disposal.

Professional Escalation Criteria

When to Seek Expert Consultation

Consult a molecular biology specialist or clinical laboratory director when:

• Assay performance falls outside established quality control parameters
• Unexpected results cannot be explained by troubleshooting
• New assays require validation before clinical use
• Regulatory compliance questions arise

When to Refer to Reference Laboratory

Refer samples to a reference laboratory when:

• Specialized equipment or expertise is required
• Confirmatory testing is needed for critical results
• Rare or emerging pathogens require identification
• Quantitative results fall outside assay range

Limitations of PCR Technology

Inherent Constraints

PCR cannot distinguish between live and dead organisms because DNA persists after cell death. RNA-based detection using reverse transcription PCR can indicate viable organisms but is subject to RNA degradation. PCR detects nucleic acid sequences, not infectious particles.

Quantitative Limitations

Conventional PCR provides only qualitative results. qPCR requires careful standardization for accurate quantification. Digital PCR has limited dynamic range. All PCR methods are subject to variation in amplification efficiency.

Interpretation Challenges

Positive results may reflect contamination, past infection, or asymptomatic carriage. Negative results do not rule out infection if inhibitors are present or if the target sequence has mutated. Clinical correlation is essential for diagnostic interpretation.

Practical Decision Framework for PCR Technology Selection

Selecting the appropriate PCR technology for a given application requires systematic evaluation of analytical requirements, sample characteristics, available resources, and regulatory constraints. The following framework provides a structured approach to technology selection based on documented performance characteristics and practical limitations.

Step 1: Define the Analytical Question

Begin by specifying whether the analysis requires qualitative detection, relative quantification, or absolute quantification. Conventional end-point PCR provides only presence-or-absence results and is suitable for genotyping, cloning, or initial screening where quantification is unnecessary. Real-time quantitative PCR (qPCR) enables relative or absolute quantification using standard curves and is appropriate for gene expression analysis, pathogen load monitoring, and viral load assessment. Digital PCR (dPCR) provides absolute quantification without standard curves and is preferred for rare mutation detection, copy number variation analysis, and applications where precise quantification at low concentrations is critical.

The choice between qPCR and dPCR depends on the required dynamic range and precision. qPCR typically spans 6-8 logs of dynamic range, while dPCR covers approximately 4-5 logs. For samples with target concentrations spanning a wide range, qPCR may be more practical. For applications requiring detection of targets present at less than 1% of total DNA, dPCR offers superior precision.

Step 2: Assess Sample Characteristics

Sample type and quality directly influence PCR technology selection. Blood samples, tissue biopsies, environmental samples, and ancient or degraded materials each present distinct challenges. For blood donor screening, automated nucleic acid amplification technology (NAT) platforms have been developed that include sample preparation, reducing costs and complexity while enabling testing of pools of up to 96 samples for hepatitis C virus, hepatitis B virus, and human immunodeficiency virus-1 [7]. More than 60 million donations per year are NAT tested worldwide using these platforms [7].

For environmental DNA samples, such as those used in fish metabarcoding, universal PCR primers targeting the 12S rRNA gene have been designed using aligned whole mitochondrial genome sequences from 880 species [8]. These primers amplify a hypervariable region of 163-185 base pairs, which is sufficiently short to work with degraded DNA [8]. When working with ancient or degraded samples, amplicon length should be kept under 200 base pairs to maximize amplification success.

Samples containing inhibitors such as heme, humic acids, or ethanol may require dilution, inhibitor-resistant polymerases, or alternative purification methods. Digital PCR is less affected by PCR inhibition than qPCR because the partitioning process dilutes inhibitors across thousands of reactions, reducing their effective concentration.

Step 3: Evaluate Quantification Requirements

Determine whether absolute quantification is necessary or whether relative quantification is sufficient. Absolute quantification requires a standard curve for qPCR or partitioning for dPCR. The 1copy BCR::ABL1 qPCR assay for monitoring chronic myeloid leukemia demonstrated linearity across a 5-log range from 0.02% to 85% on the international scale, with a coefficient of determination of 0.9949 [12]. Coefficients of variation at 14.1% and 0.233% on the international scale were 14.4% and 15.6%, respectively [12]. These performance characteristics should be considered when establishing acceptable precision limits for clinical monitoring.

For applications requiring detection at clinically critical thresholds, concordance between methods should be evaluated. The 1copy BCR::ABL1 qPCR assay showed 98.6% concordance with an established assay at the greater than 10% international scale threshold, 100% at greater than 1%, and 95.9% at less than or equal to 0.1% [12]. When selecting a technology for clinical use, verify that published concordance data meet the required diagnostic accuracy.

Step 4: Consider Throughput and Cost Constraints

High-throughput PCR systems are suitable for large-scale screening applications such as blood donor testing, where automation has rendered possible the reduction of pool size in conjunction with increased throughput and sensitivity [7]. Microfluidics-based PCR devices for point-of-care applications offer portability and rapid turnaround time but may have lower throughput [3].

Digital PCR has higher per-sample cost than qPCR due to specialized consumables and instrumentation. The limited dynamic range of dPCR may require multiple dilutions for samples with unknown target concentrations, increasing cost and hands-on time. For routine quantification with wide dynamic range, qPCR remains more cost-effective.

Step 5: Verify Regulatory and Quality Requirements

Clinical diagnostic applications require validation according to established guidelines. Precision, linearity, and detection capability should be assessed according to Clinical and Laboratory Standards Institute guidelines [12]. Limit of blank, limit of detection, and limit of quantification should be determined for each assay. For blood donor screening, regulatory requirements may specify minimum sensitivity and specificity thresholds.

When implementing a new PCR technology, include appropriate controls in every run: positive controls to verify reagent function, negative controls to detect contamination, and internal positive controls to identify inhibition. Document all quality control results and establish acceptable ranges for control values.

Records and Measurements

Maintain the following records for each PCR assay:

• Assay name and version
• Template source and extraction method
• Primer and probe sequences and concentrations
• Thermal cycling parameters
• Instrument used and calibration date
• Control results (positive, negative, internal)
• Sample results with Ct values or copy numbers
• Standard curve parameters (slope, efficiency, R-squared) for qPCR
• Number of partitions and positive partitions for dPCR
• Any deviations from standard protocol

For qPCR, record amplification efficiency calculated from the standard curve slope. Acceptable efficiency ranges from 90% to 110%, corresponding to slopes between -3.6 and -3.1. Record the coefficient of determination (R-squared) for the standard curve, values above 0.98 indicate acceptable linearity.

For dPCR, record the total number of partitions analyzed and the number of positive partitions. Calculate target concentration using Poisson statistics and record the confidence interval. Document any partitions excluded due to quality metrics.

Common Failure Patterns in Technology Selection

Overestimating dynamic range requirements. Selecting dPCR when qPCR would suffice increases cost without proportional benefit. If target concentrations are expected to span more than 4 logs, qPCR is more practical.

Underestimating inhibition effects. Samples containing inhibitors may produce false-negative results with qPCR. Digital PCR may mitigate this issue, but inhibition can still reduce partition fluorescence. Include internal positive controls to detect inhibition in both technologies.

Ignoring allelic dropout. Primer binding site polymorphisms can cause preferential amplification of one allele, leading to misclassification of heterozygous samples. In white-tailed deer prion protein gene sequencing, a widely used forward primer caused allelic dropout that misclassified 128 out of 586 deer previously classified as homozygous [16]. Redesigning primers to avoid polymorphic sites corrected this error [16]. When designing primers for polymorphic targets, verify primer binding sites against known sequence variants.

Neglecting standardization. For clinical applications, results must be reported on an international scale when available. The BCR::ABL1 qPCR assay comparison study demonstrated the importance of using standardized reporting units to enable comparison across laboratories and methods [12].

Welfare and Safety Context

PCR technology selection has direct implications for patient and population welfare. In blood donor screening, the introduction of NAT testing reduced the residual risk of virus transmission by blood components and products to almost zero [7]. When severe acute respiratory syndrome coronavirus and West Nile virus emerged, NAT enabled manufacturers and transfusion centres to instantly introduce sensitive and specific screening tests [7]. Selecting the appropriate PCR technology for emerging pathogens can accelerate diagnostic capacity and improve public health response.

In wildlife disease surveillance, PCR-based genotyping can map susceptibility to chronic wasting disease across the landscape, focusing on prion protein variant frequencies to improve epidemiological risk models [16]. Accurate genotyping requires careful primer design to avoid allelic dropout, as misclassification can lead to incorrect risk assessments.

Laboratory safety must be considered when selecting PCR technology. The World Health Organization Laboratory Biosafety Manual provides guidance for safe handling of infectious materials [1]. Work with known pathogens requires containment facilities and practices appropriate to the biosafety level. Digital PCR may reduce contamination risk because the sample is partitioned into sealed reactions, but standard biosafety precautions still apply.

Professional Escalation Criteria

Consult a molecular biology specialist or clinical laboratory director when:

• Assay performance consistently falls outside established quality control parameters
• Unexpected results cannot be explained by troubleshooting
• New assays require validation before clinical use
• Regulatory compliance questions arise regarding technology selection
• Rare or emerging pathogens require identification using non-standard methods

Refer samples to a reference laboratory when:

• Specialized equipment or expertise is required for the selected technology
• Confirmatory testing is needed for critical clinical results
• Quantitative results fall outside the validated assay range
• Discordant results between different PCR technologies require resolution

Frequently Asked Questions

What is the difference between PCR and qPCR?

Conventional PCR detects amplified products at the end of the reaction, providing qualitative results. Real-time quantitative PCR (qPCR) monitors amplification during each cycle using fluorescent detection, allowing quantification of starting template. qPCR provides cycle threshold values that correlate with target concentration.

How does digital PCR differ from qPCR?

Digital PCR partitions the sample into thousands of individual reactions and counts positive partitions to calculate absolute target concentration without requiring standard curves. qPCR relies on standard curves for quantification and has a wider dynamic range. Digital PCR offers higher precision for rare target detection.

What are the main applications of PCR in diagnostics?

PCR is used for infectious disease diagnosis, genetic disorder screening, cancer monitoring, and forensic identification. Real-time reverse transcription PCR is the gold standard for SARS-CoV-2 diagnosis [3]. NAT testing screens blood donations for HIV, hepatitis viruses, and emerging pathogens [7].

How do I choose between qPCR and digital PCR?

Choose qPCR for routine quantification with wide dynamic range, gene expression analysis, and viral load monitoring. Choose digital PCR for absolute quantification without standards, rare mutation detection, and copy number variation analysis. Consider cost, throughput, and required precision.

What causes PCR failure?

Common causes include degraded template, inhibitors in the sample, incorrect primer design, suboptimal annealing temperature, inactive polymerase, and contamination. Systematic troubleshooting should address each potential cause. Include appropriate controls to identify the source of failure.

How can I prevent PCR contamination?

Use separate areas for pre- and post-amplification work. Use dedicated pipettes with aerosol-resistant tips. Include no-template controls in every run. Decontaminate surfaces and equipment regularly. Use uracil-DNA glycosylase to prevent carryover contamination.

What is the role of PCR in blood donor screening?

NAT testing for blood donor screening detects viral nucleic acids, reducing the window period between infection and detectable markers. More than 60 million donations per year are NAT tested worldwide, reducing residual risk of virus transmission to almost zero [7].

Can PCR detect all pathogens?

PCR detects only the specific target sequences included in the assay design. It cannot detect unknown or unexpected pathogens. Multiplex PCR can detect multiple targets simultaneously, but comprehensive pathogen detection requires broad-range assays or sequencing approaches.

Related Guides

• Digital Droplet PCR vs qPCR: A Practical Comparison for Absolute Quantification
• Digital Droplet PCR (ddPCR) for Absolute Quantification of Viral Load in Veterinary Diagnostics: Principles and Applications
• Digital Droplet PCR (ddPCR) for Absolute Quantification of Viral Load in Veterinary Pathogens: Principles and Applications
• Development and Evaluation of a Digital PCR Assay for the Detection of African Swine Fever Virus in Oral Fluids
• Digital Droplet PCR for Absolute Quantification of Feline Enteric Coronavirus RNA in Fecal Samples: Diagnostic Utility and Prognostic Implications

References and Further Reading

• Laboratory Biosafety Manual. World Health Organization.
• NCBI Literature Resources. National Center for Biotechnology Information.
• PCR past, present and future.. BioTechniques, 2020.
• The Evolution of Next-Generation Sequencing Technologies.. Methods in molecular biology (Clifton, N.J.), 2025.
• Viral Pneumonia.. 2026.
• Keratoconjunctivitis.. 2026.
• History and Future of Nucleic Acid Amplification Technology Blood Donor Testing.. Transfusion medicine and hemotherapy : offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie, 2019.
• MiFish, a set of universal PCR primers for metabarcoding environmental DNA from fishes: detection of more than 230 subtropical marine species.. Royal Society open science, 2015.
• Diagnosing past infections: A review.. Diagnostic microbiology and infectious disease, 2025.
• The evolution of next-generation sequencing technologies.. ArXiv, 2023.
• Comparative plastid genome analysis of four different variants of Gastrodia elata and development of molecular markers based on plastid DNA sequences.. 2026.
• Analytical performance evaluation and method comparison of the 1copy BCR::ABL1 qPCR assay for monitoring chronic myeloid leukemia.. 2026.
• Pathogenic germline variants associated with different HER2 expression among breast cancer patients.. 2026.
• Large-scale gastroenteritis outbreaks occurring in a university town of Guangzhou city during the global wave of the norovirus in China.. 2026.
• Mapping Current and Emerging Laboratory Techniques for Haemoglobinopathy Carrier Detection and Prevention: A Narrative Review from the HELIOS Action.. 2026.
• Odocoileus virginianus PRNP sequencing reveals AF (Q&lt,sub&gt,95&lt,/sub&gt,G&lt,sub&gt,96&lt,/sub&gt,/H&lt,sub&gt,95&lt,/sub&gt,G&lt,sub&gt,96&lt,/sub&gt,) advantage over AC (Q&lt,sub&gt,95&lt,/sub&gt,G&lt,sub&gt,96&lt,/sub&gt,/Q&lt,sub&gt,95&lt,/sub&gt,S&lt,sub&gt,96&lt,/sub&gt,) against chronic wasting disease.. 2026.
• Analysis of the Advantages and Disadvantages of PCR Technology and Ideas for Improvement. Theoretical and Natural Science, 2025.
• Brief Introduction to PCR Technology and Its Application. 2002.
• [PCR-derived technology in gene identification and typing of Yersinia pestis].. Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine], 2015.
• Real-time PCR, History and Fluorgenic Chemistries. 2007.
• PCR-derived technology in gene identification and typing of Yersinia pestis. Zhonghua Yu Fang Yi Xue Za Zhi Chinese Journal of Preventive Medicine, 2015.
• Digital PCR and its application in biological detection. Yi Chuan Hereditas, 2020.
• Application of digital PCR technology in the precise molecular diagnosis: opportunities and challenges. Chinese Journal of Laboratory Medicine, 2022.

This article is educational and does not replace validated laboratory procedures, institutional biosafety review, manufacturer instructions, or professional interpretation.