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Paper Information

Journal:   IRANIAN JOURNAL OF PUBLIC HEALTH   AUGUST 2014 , Volume 43 , Number SUPPLEMENT 2; Page(s) 71 To 71.
 
Paper: 

DEVELOPMENT AND CLINICAL VALIDATION OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION METHOD FOR RAPID DETECTION OF BORDETELLA PERTUSSIS BASED ON INSERTION SEQUENCE IS481

 
 
Author(s):  ALLAHYAR TORKAMAN MOHAMMAD REZA*, NIKBIN VAJIHE SADAT, NAKHOST LOTFI MASOUMEH, SHAHCHERAGHI FERESHTEH
 
* DEPARTMENT OF MICROBIOLOGY, MIC RESEARCH CENTER, PASTEUR INSTITUTE OF IRAN, TEHRAN, IRAN
 
Abstract: 

Background: In spite of the existence of conventional culture method and developing PCR-based assays, yet the accurate detection of Bordetella pertussis is a problem. In this study, we developed a Loop-Mediated Isothermal Amplification (LAMP) assay based on IS481 and it has evaluated the efficiency, rapidity, sensitivity, and specificity of IS481 LAMP in comparison with real-time PCR with the same target for the detection of B. pertussis clinical specimens.
Methods: One hundred and sixty-five nasopharyngeal swabs were used to be assessed by LAMP and real-time PCR. Amplified LAMP products were detected by adding SYBRR Safe DNA Gel Stain. Illumination under UV light and the green color for positive samples were visible by unarmed eyes.
Results: The sensitivity and specificity of the LAMP assay were reasonable in comparison with real-time PCR. In this study, they were 87.5% and 92.9% for IS481 target, respectively. Amplification of 10 fg by IS481-LAMP showed the sensitivity of 2.26 genomic copies of B. pertussis.
Conclusion: The developed LAMP assay based on IS481 target helps rapid and sensitive detection of B. pertussis, and it is cheaper than previous conventional real-time PCR. This IS-481 LAMP assay has the merit of being used as a diagnostic method for B. pertussis.

 
Keyword(s): LAMP, BORDETELLA PERTUSSIS, IS481
 
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