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35 Differences Between PCR and ELISA Assays in Laboratory Testing

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Lab Wale

On December 1, 2023

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Frequently Asked Questions (FAQ’S)

Enzyme-Linked Immunosorbent Assay (ELISA) and Polymerase Chain Reaction (PCR) are two crucial methods used in laboratory testing for various objectives.

A laboratory method used in molecular biology, PCR stands for Polymerase Chain Reaction and is used to amplify and analyze DNA (deoxyribonucleic acid). A potent and popular technique for identifying, measuring, and analyzing certain DNA sequences is PCR. Kary Mullis, who received the 1993 Nobel Prize in Chemistry for developing the process initially, did so in the 1980s.

The versatility and accuracy of PCR have made it a vital instrument in molecular biology, significantly advancing scientific fields such as genetics, medicine, and other fields.

Enzyme-Linked Immunosorbent Assay is referred to as ELISA. It is a popular laboratory method for identifying and measuring chemicals in biological samples, including proteins, antibodies, hormones, and other compounds. Medical diagnostics, research, and quality control procedures frequently use ELISA.

Using certain antibodies that have the ability to attach to the target molecule of interest is the fundamental idea behind ELISA.

There are various ELISA variants, including sandwich, direct, indirect, and competitive ELISA, each with specialized uses based on the kind of molecule being found and the objectives of the experiment. Clinical laboratories employ ELISA extensively for disease diagnosis, treatment response monitoring, and a variety of research applications in immunology, microbiology, and biochemistry.

S.No.	Aspects	PCR	ELISA
1	Full Name	Polymerase Chain Reaction	Enzyme-Linked Immunosorbent Assay
2	Type of Assay	Nucleic Acid Amplification	Protein Detection
3	Purpose	Detecting DNA/RNA	Detecting Proteins
4	Sensitivity	High sensitivity	Moderate sensitivity
5	Specificity	High specificity	Moderate specificity
6	Sample Type	DNA/RNA samples	Serum, plasma, or other biological fluids
7	Detection Target	Genetic sequences	Antigens or antibodies
8	Amplification Step	Yes	No
9	Enzymes Used	DNA polymerase	Enzymes (e.g., horseradish peroxidase)
10	Endpoint Detection	Real-time monitoring or gel electrophoresis	Colorimetric or fluorescent signals
11	Quantification	Quantitative and qualitative results	Quantitative or qualitative results
12	Time Required	Several hours to a few days	Hours
13	Cost	Expensive	Relatively Inexpensive
14	Automation	Semi-automated to fully automated systems	Typically manual or semi-automated
15	Multiplexing	Possible with multiplex PCR	Limited multiplexing capabilities
16	Analyte Concentration	Low concentrations of nucleic acids	Higher concentrations of antigens/antibodies
17	Detection Method	Fluorescence or gel electrophoresis	Colorimetric or chemiluminescence
18	Clinical Applications	Diagnosing infectious diseases, genetic disorders	Detecting antibodies, antigen presence
19	Sensitivity to Contaminants	Susceptible to contamination	Less susceptible to contamination
20	Sample Storage	Requires cold storage for stability	Samples can be stored at room temperature
21	False Positives	Less common	More common
22	False Negatives	Less common	Less common
23	Throughput	Lower throughput	Higher throughput
24	Test Timeframe	Longer turnaround time	Faster results
25	Sensitivity to Inhibitors	Sensitive to PCR inhibitors	Less sensitive to inhibitors
26	Clinical Validation	Often requires confirmatory tests	May not require confirmatory tests
27	Target Molecule Stability	DNA/RNA can be stable for a long time	Proteins may degrade over time
28	Detection Mechanism	Amplification of target sequences	Antigen-antibody interactions
29	Amplification Cycles	Multiple amplification cycles are required	No amplification cycles
30	Equipment	PCR machine or thermocycler	ELISA plate reader and washer
31	Skill Level Required	Requires skilled technicians	Less technical expertise required
32	Sample Volume	Small sample volumes required	Larger sample volumes required
33	Variability in Results	Less variability	More variability
34	Quantitative Range	Wide quantitative range	Limited quantitative range
35	Historical Use	Common in molecular biology and genetics	Common in immunology and serology

Frequently Asked Questions (FAQ’S)

Q1. What makes RT-PCR different from PCR?

While RT-PCR (Reverse Transcription Polymerase Chain Reaction) is used to amplify RNA, PCR (Polymerase Chain Reaction) is used to amplify DNA. Reverse transcription is a step in RT-PCR that turns RNA into complementary DNA (cDNA).

Q2. What function do primers serve in PCR?

During PCR, primers are brief DNA sequences that attach to the target DNA area and act as the beginning sites for DNA polymerase to synthesize new DNA.

Q3. How does PCR function?

The three primary processes of PCR are extension, annealing, and denaturation. The process of making a copy of the original DNA involves heating DNA to separate its strands, primers binding to the target sequence, and DNA polymerase extending the new strand.

Q4. Which ELISA kinds are there?

ELISA comes in a variety of forms, such as competitive, sandwich, direct, and indirect. According to the target molecule and the required assay sensitivity, each kind has particular applications.

Q5. What benefit does ELISA offer in terms of diagnostics?

ELISA has the ability to analyze many samples at once and is very specific and sensitive. It is extensively employed in clinical diagnostics to identify different biomarkers, autoimmune conditions, and infectious infections.

Q6. Is ELISA automatable?

Indeed, automation of ELISA can improve its performance and lower assay variability. High-throughput laboratories frequently use automated ELISA systems.

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35 Differences Between PCR and ELISA Assays in Laboratory Testing

Frequently Asked Questions (FAQ’S)

Q1. What makes RT-PCR different from PCR?