Associated technologies ensures complete loop mediated isothermal amplification platform for pathogen diagnosis

  • Selvam M Masilamani
  • Divya Sasitharan
  • Paul W Manohar

Abstract

Loop Mediated Isothermal Amplification (LAMP) assay could be a useful adjunct diagnostic assay along with the conventional methods that would preclude the requirement of continuous maintenance of pure cultures. Moreover, LAMP assay is simple, rapid, specific and sensitive for the detection of pathogens. Having developed and validated LAMP method for the detection of an isolated pathogen, Escherichia coli O157:H7, an attempt was made to progress the LAMP platform to realistic point of care for resource-poor endemic areas. Reaction time of the LAMP method was only 1 h and also, the amplification products of O157, which had the corresponding target genes, turned green by visual inspection when added with Calcein/Sybr green. However, sample preparation and lyophilized master mix preparation before LAMP assay as well as developing a closed detection system for detection of LAMP amplified products remained a quest. Hence, the current study was conducted to develop a lyophilized LAMP master mix for easy platform to take LAMP to realistic point of care and Dot-Elisa based ‘lateral flow dipstick’ that could ease the detection of LAMP products in a closed environment. Sample preparation is another associated technology that is yet-to-be developed.

Keywords: Loop mediated isothermal amplification assay, polymerase chain reaction, lyophilized master mix, and closed amplification system.

African Journal of Biotechnology Vol. 12(42), pp. 6049-6054

Author Biographies

Selvam M Masilamani
Department of Biotechnology, Sathyabama University, Chennai, 600119 India
Divya Sasitharan
Department of Biotechnology, Sathyabama University, Chennai, 600119 India
Paul W Manohar
Department of Veterinary Microbiology, Madras Veterinary University, Chennai, 600007, India.
Published
2016-06-06
Section
Articles

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eISSN: 1684-5315