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Updated: Sep 8, 2025
Author Spotlight: Addressing Regulatory Gaps in Molecular Studies by Quantifying Viral Vectors in Complex Matrices
Published on: July 14, 2023
Preliminary evaluation of inactivated antigens as diagnostic controls in molecular detection of vector-borne diseases
Manjini Harshavarthini1, Subbiah Kombiah1,2, Satyendra Pandey1
1ICMR-National Institute of Research in Tribal Health, Jabalpur, India.
Background Objectives:
Molecular tests play a crucial role in diagnosis, however, acquiring appropriate controls is challenging owing to strict biosafety and biosecurity regulations. In this work, we suggest an alternative to live virus for inclusion as positive controls in molecular testing.
Methods:
We used inactivated antigens provided with ELISA kits, both as they are and after spiking the vector samples. The extracted RNA/DNA was used as a positive control in RT-PCR/PCR reactions.
Results:
The PCR product of expected size and sequencing results confirmed that the inactivated antigens can be used as positive controls for dengue, chikungunya, Japanese encephalitis and lymphatic filariasis in molecular tests.
Interpretation Conclusion:
We demonstrate that the inactivated antigens supplied with ELISA kits or obtained from microscopic slides can be used as positive controls in molecular tests while testing human and vector samples. This will be a useful technique for resource-constrained laboratories and during emergencies.
Insights
Inactivated antigens from ELISA kits or slides can serve as reliable positive controls for molecular tests, overcoming challenges with live virus acquisition. This method is valuable for resource-limited settings and emergency diagnostics.
Area of Science:
- Medical diagnostics
- Molecular biology
- Virology
Background:
- Molecular tests are vital for disease diagnosis.
- Acquiring live virus controls for molecular testing is difficult due to strict biosafety regulations.
- An alternative to live virus positive controls is needed.
Purpose of the Study:
- To evaluate inactivated antigens as positive controls for molecular testing.
- To provide a viable alternative to live viruses for molecular diagnostic controls.
Main Methods:
- Inactivated antigens from ELISA kits and microscopic slides were used.
- Antigens were tested directly and after spiking vector samples.
- Extracted RNA/DNA from inactivated antigens served as positive controls in RT-PCR/PCR.
Main Results:
- PCR products of the expected size confirmed successful amplification.
- Sequencing validated the use of inactivated antigens as controls.
- Effective as positive controls for Dengue, Chikungunya, Japanese Encephalitis, and Lymphatic Filariasis molecular tests.
Conclusions:
- Inactivated antigens are suitable positive controls for molecular tests on human and vector samples.
- This technique offers a practical solution for laboratories in resource-constrained settings.
- The method is particularly useful during public health emergencies.
