Development and evaluation of HIV-۱ diagnostic kits based on recombinant reversetranscription loop-mediated isothermal amplification assay

سال انتشار: 1402
محل انتشار: اولین کنفرانس بین المللی علم و فناوری خوارزمی
کد COI اختصاصی: KHWARIZ01_105
زبان مقاله: انگلیسی
تعداد مشاهده: 178

نویسندگان

Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol,Iran.

Fatemeh Tohidi

Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol,Iran.bBiomedical and Microbial Advanced Technologies (BMAT) Research Center, Health Research Institute, Babol University ofMedical Scien

Hosein Shahsavarani

Dept. of Cell and Molecular Biology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran.

چکیده

The identification of HIV-۱ at the point-of-care or within resource-constrained settings presents asignificant challenge due to limitations in time and cost. Currently, PCR and real-time PCR tests stand as the soledependable method for diagnosing recent infections during the window period following infection and prior toseroconversion. Nonetheless, these tests are highly time-consuming and costly, necessitating expensivematerials, a thermal cycler, gel electrophoresis, or computational and data analysis software, all of which areexclusively suitable for well-equipped laboratory environments. Given the gravity of the disease, there exists anurgent need for a straightforward, economical, rapid, and accessible diagnostic approach. The ReverseTranscription Loop-Mediated Isothermal Amplification (RT-LAMP) technique offers a distinct molecular and costeffectivealternative method with exceptional specificity and sensitivity for the creation of a swift nucleic acidamplification test (NAAT). This amplification is conducted in a tube containing a blend of primers, reversetranscriptase enzyme, and Bst DNA polymerase at a temperature of ۶۰ °C for ۶۰ minutes. Furthermore, a basicwater bath or heat block suffices for the amplification of the LAMP products, culminating in direct visualdetection facilitated by the addition of a slight amount of Sybr green I fluorescent dye to the microtube underUV light. In this research, we initially fine-tuned the expression of recombinant Bst polymerase and subsequentlycarried out the RT-LAMP technique using six specific primers targeting the IN coding region of the POL gene torecognize different HIV-۱ subtypes. As a result, we foresee that the outcomes of this study will pave the way forprogress towards the development and production of an HIV-۱ diagnostic kit in Iran in the near future.

کلیدواژه ها

HIV-۱; RT-LAMP; Recombinant Bst DNA Polymerase

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