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Enzymatic amplification of lactate dehydrogenase-elevating virus
J T van der Logt1, J Kissing, W J Melchers
1Department of Medical Microbiology, University of Nijmegen, The Netherlands.
Journal of Clinical Microbiology
|August 1, 1994
Summary
A new PCR assay effectively detects lactate dehydrogenase-elevating virus (LDV) in mice, offering a more reliable diagnostic tool than the traditional LDH enzyme assay. This method shows high specificity and stability, even with sample degradation.
Area of Science:
- Virology
- Molecular Biology
- Diagnostic Assays
Background:
- Lactate dehydrogenase-elevating virus (LDV) is an important pathogen in mouse models.
- Current diagnostic methods for LDV, such as the lactate dehydrogenase (LDH) enzyme assay, have limitations.
Purpose of the Study:
- To develop and validate a highly specific and sensitive PCR assay for LDV detection.
- To compare the efficacy of the new PCR assay with the conventional LDH enzyme assay.
Main Methods:
- Development of a PCR assay using primers targeting the ORF7 gene (nucleocapsid protein VP1) of LDV.
- Testing the specificity of the PCR assay against common murine viruses and related arteriviruses.
- Detection of LDV in plasma samples from experimentally infected mice during acute and persistent phases.
- Evaluation of PCR assay performance with contaminated Plasmodium berghei parasite pools.
- Comparison of PCR results with LDH enzyme assay results.
- Assessment of sample stability under various storage conditions.
Main Results:
- The developed PCR assay demonstrated high specificity, with no cross-reactivity with other murine viruses or related arteriviruses.
- LDV was reliably detected in mouse plasma during both acute and persistent infection phases.
- The PCR assay successfully identified LDV in contaminated parasite pools and in infected mice.
- PCR assay results showed complete agreement with LDH enzyme assay findings.
- The PCR assay did not produce false-positive results with hemolyzed samples, unlike the LDH assay.
- Plasma sample integrity was maintained for LDV detection by PCR after storage up to 8 days at various temperatures.
Conclusions:
- The developed PCR assay is a sensitive, specific, and robust diagnostic tool for LDV detection.
- The PCR assay offers advantages over the LDH enzyme assay, including improved specificity and resistance to sample degradation.
- This PCR technique has the potential to replace the LDH test for routine LDV diagnostics.