Paper Details

Development of a loop-mediated isothermal amplification of DNA by using florescent probe in detection of salmonella

Bita Bagheri, Ali Karami, Gita Bagheri

DOI: https://dx.doi.org/10.12692/ijb/6.2.6-14

Int. J. Biosci. 6(2), 6-14. January, 2015. (PDF)

Abstract:

Salmonellas are a group of Entrobacteriace that cause Infection disease in human and animals .Thyphoid, Bacteriemi ,Entrocolit and Salmonelose are cause by this bacteria turned to be a major health problem in developing countries ,there for quick , accurate ,reliable and on time diagnosis seem to be vital to prevent it from spreading widely and become epidemic . Different method are used to diagnosing of salmonellas, all of this method require long time to diagnose, numerous bacteria ,expensive and specific lab equipment and expert lab personal. In this research, we used lamp (loop mediated Isothermal amplification of DNA) method with florescent probe to diagnosis five types of salmonella and compare with PCR and lamp. Then optimized this method by the time, probe concentration, and temperature and optimum condition was obtained at temperature on LAMP using fluorescent probe at 37°C, time at 0.2 and concentration of pobe at 4%. According to the obtained results rapid ,accurate ,economic, easy to use , specific and sensitive are the most important factors to evaluate and assay to detect of this method ( LAMP by using florescent probe ) have extremely all of them that was be used in detection of pathogenic viruses, bactereria, parasites and other medical diagnosis laboratories ,lea gal medical ,agricultural and research filed that takes tie time lees than PCR and LAMP ,this method is independent to cycles temperature and thermo cycling that replacement with thermo block and color detection is more accurate and facilitate than turbidity detection in LAMP.