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Published on: March 13, 2018
Multiplex Loop-Mediated Isothermal Amplification Coupled Lateral Flow Assay for Point-of-Care Detection of Syphilis
Krishnamoorthy Priya1, Elangovan Saranya1, Ashish Kapoor2
1Department of Genetic Engineering, School of Bioengineering, Faculty of Engineering and Technology, SRM Institute of Science and Technology, SRM Nagar, Kattankulathur, Kanchipuram, Chennai, Tamil Nadu 603203 India.
A new multiplex loop-mediated isothermal amplification coupled lateral flow assay (multiplex LAMP-LFA) offers rapid and sensitive syphilis detection. This point-of-care test shows high accuracy, addressing limitations of current diagnostic methods.
Area of Science:
- Infectious Diseases
- Molecular Diagnostics
- Point-of-Care Testing
Background:
- Syphilis, caused by *T. pallidum*, is a re-emerging global health concern with over 5 million annual cases.
- Current serological diagnostic methods lack sensitivity in early infection stages and have other limitations.
- There is a critical need for rapid, simple, sensitive, and cost-effective point-of-care diagnostic tools for syphilis.
Purpose of the Study:
- To develop a novel multiplex loop-mediated isothermal amplification coupled lateral flow assay (multiplex LAMP-LFA) for syphilis detection.
- To evaluate the sensitivity, specificity, and speed of the developed multiplex LAMP-LFA assay.
Main Methods:
- Developed a multiplex LAMP assay to amplify two genes: *polA* (target) and human *RNase P* (internal control).
- Utilized lateral flow assay (LFA) strips with Anti-FITC and Anti-DIG antibodies for rapid detection of amplified products.
- Assessed the assay's performance using defined copy numbers and tested with 130 clinically suspected syphilis cases and 50 healthy individuals.
Main Results:
- The multiplex LAMP-LFA achieved a detection limit of 3.8 × 103 copies/mL with high specificity.
- In clinical evaluations, the assay demonstrated a sensitivity of 93.84% and a specificity of 100%.
- Results were obtained within 5 minutes of sample application on the LFA strip.
Conclusions:
- The developed multiplex LAMP-LFA is a sensitive, rapid, and visual method for detecting *T. pallidum*.
- This assay overcomes limitations of traditional Non-Treponemal and Treponemal tests, offering an affordable alternative.
- Multiplex LAMP-LFA holds significant potential for point-of-care syphilis diagnostics, especially in resource-limited settings.
