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   improved visual detection of speb gene in streptococcus pyogenes isolates by real-time loop-mediated isothermal amplification turbidimetry method  
   
نویسنده simon azi simon ,subbiah suresh kumar ,than leslie thian lung ,osman malina ,hamat rukman awang
منبع jundishapur journal of microbiology - 2021 - دوره : 14 - شماره : 2 - صفحه:1 -7
چکیده    Background: groupa streptococcus (gas) causes a wide array of clinical manifestations ranging from mild pharyngitis to suppurative and non-suppurative severe debilitating diseases. hence, a simple, rapid detection method with high sensitivity and specificity is needed. objectives: this study embarked on the visual detection of the streptococcal pyrogenic exotoxin b (speb) gene by real-time turbidimetry and loop-mediated isothermal amplification (rt-lamp) methods. the real-time monitoring of the sigmoidal graph generated from a turbidimetry method was incorporated in the assay. methods: the amplification of the speb gene was virtually observed in real-time monitoring of the graph (sigmoidal curve) generated via a turbidimeter, thus providing a “guide” to accurately estimate the time to positivity for the gene detection. results: the targeted gene was detected at 15 min but was optimally amplified within 45 min at an isothermal temperature of 63°c with 100% specificity using an established set of primers. the formation of sigmoidal curves was correlated with other visual observations by the naked eye (from orange to green), ultra-violet light (green fluorescence), and agarose gel electrophoresis. the improved detection limit of the real-time rt-lamp assay was also observed compared to conventional pcr assay (0.001 pg/μl versus 1 ng/μl). conclusions: the improved visual detection of rt-lamp assay could provide additional insight for rapid, cost-effective, and reliable identification of gas via speb gene detection in low or middle-income countries. it could also be a very important tool to improve the healthcare management of patients infected with gas in the future.
کلیدواژه rapid detection ,real-time loop-mediated isothermal amplification ,point-of-care testing ,streptococcal pyrogenic exotoxin b
آدرس ebonyi state university, faculty of health sciences and technology, department of medical laboratory science, nigeria, universiti putra malaysia, faculty of medicine and health sciences, department of medical microbiology, malaysia, universiti putra malaysia, faculty of medicine and health sciences, department of medical microbiology, malaysia, universiti putra malaysia, faculty of medicine and health sciences, department of medical microbiology, malaysia, universiti putra malaysia, faculty of medicine and health sciences, department of medical microbiology, malaysia
پست الکترونیکی rukman@upm.edu.my
 
     
   
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