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In Association with
A comparative study of the performance of loop mediated isothermal amplification (LAMP) and PCR for diagnosis of gonorrhea
6th World Congress on Biotechnology
Shazia Shaheen Mir, Asif Anas, Tanzeel, Nimisha Patel, Uzair Ahmad and Arif Ali
Jamia Millia Islamia, India
Posters-Accepted Abstracts: J Biotechnol Biomater
Abstract
Conventional methods for detection of Neisseria gonorrhoeae causing gonorrhea have been gradually replaced by nucleic acid amplification tests (NAATs). However, all these NAATs have inherent disadvantages of requiring a high precision instrument for amplification or an elaborate complicated method for detection of amplified products. Thus, for settings with minimal facilities, there is a need for a simple reliable test that would permit the screening of N. gonorrhoeae. Therefore, in this study we develop and compare LAMP with PCR for detection of N. gonorrhoeae. LAMP has been reported to amplify DNA without any special requirement with high specificity, efficiency and rapidity under isothermal conditions. LAMP assay and PCR were developed with self-designed primers for opa gene separately. LAMP and PCR were standardized by using N. gonorrhoeae (ATCC 49226). Serial dilutions of DNA extracted from N. gonorrhoeae (ATCC 49226) were used to determine the sensitivity of the assay. Specificity was checked using N. gonorrhoeae (ATCC 49226, clinical isolates n=41), N. sicca (ATCC 29193, clinical isolates n=6), N. meningitidis (ATCC 13078, clinical isolates n=2), Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 25923), Pseudomonas aeruginosa (ATCC 27853), Streptococcus pyogenes and Chlamydia trachomatis. LAMP was highly specific for N. gonorrhoeae showing no cross-reaction. LAMP showed amplification within 30 minutes. Detection limit of LAMP and PCR were 4 fg and 4 pg respectively. LAMP showed concordance (94.4%) with PCR in clinical samples. This LAMP assay offers a highly sensitive and specific assay for the detection and confirmation of N. gonorrhoeae, thereby saving time and cost.Biography
Email: shaziamir29@gmail.com
