Abstract
Introduction. Due to the high clinical significance of herpesvirus diseases, the searching of fast and effective methods for their diagnosis remains relevant.The aim of the study was to evaluate the diagnostic efficiency of the loop-mediated isothermal amplification of DNA with real-time fluorescent detection (RT-LAMP) with SYTO-82 dye on a model of herpes simplex virus (HSV) infection.Materials and methods. A total of 44 urogenital swabs containing type 1 and type 2 HSV DNA and 43 swabs without HSV DNA, including 33 samples containing the DNA of cytomegalovirus, Epstein-Barr virus and herpesvirus type 6, were studied. For RT-LAMP, Bst 2.0 WarmStart DNA polymerase, SYTO-82 dye, LAMP primers were used.Results. The high efficiency of HSV DNA detection in the RT-LAMP reaction with SYTO-82 dye was shown. RT-LAMP in optimal conditions allowed to reduce reaction time for 2-3 times compared to real-time PCR (to 35 minutes). Analytical sensitivity of HSV type 1 and 2 detection in RT-LAMP was 103 copies of DNA/ml. The diagnostic sensitivity and specificity of the RT-LAMP diagnosis of HSV infection were 96% and 100%, respectively.Discussion. RT-LAMP method has a high sensitivity and specificity comparable to RTPCR, while the risk of false positive results obtaining is minimal.Conclusion. Thus, the reaction of RT-LAMP with SYTO-82 dye allows quickly, with high sensitivity and specificity to detect HSV DNA in clinical material and can be considered as a promising point-of-care testing method.
Highlights
Ключевые слова: петлевая изотермическая амплификация ДНК; LAMP; SYTO-82; вирус простого герпеса; диагностика на месте оказания медицинской помощи
Due to the high clinical significance of herpesvirus diseases, the searching of fast and effective methods for their diagnosis remains relevant.The aim of the study was to evaluate the diagnostic efficiency of the loop-mediated isothermal amplification of DNA with real-time fluorescent detection (RT-LAMP) with SYTO-82 dye on a model of herpes simplex virus (HSV) infection
RT-LAMP in optimal conditions allowed to reduce reaction time for 2-3 times compared to real-time PCR
Summary
Смирнова Д.И., Петруша О.А., Грачёва А.В., Волынская Е.А., Зверев В.В., Файзулоев Е.Б. Целью работы была оценка диагностической эффективности метода петлевой изотермической амплификации ДНК с флуоресцентной детекцией в режиме реального времени (LAMP-РВ) с красителем SYTO-82 на модели инфекции вирусов простого герпеса (ВПГ). Показана высокая эффективность использования красителя SYTO-82 для детекции ДНК ВПГ 1-го и 2-го типа в реакции LAMP-РВ. Реакция LAMP-РВ с красителем SYTO-82 позволяет быстро, с высокой чувствительностью и специфичностью выявлять ДНК ВПГ в клиническом материале и может рассматриваться как перспективный метод диагностики герпесвирусных инфекций на месте лечения. Быстрая диагностика генитального герпеса методом петлевой изотермической амплификации ДНК с флуоресцентной детекцией Журнал микробиологии, эпидемиологии и иммунобиологии (ЖМЭИ). Due to the high clinical significance of herpesvirus diseases, the searching of fast and effective methods for their diagnosis remains relevant.The aim of the study was to evaluate the diagnostic efficiency of the loop-mediated isothermal amplification of DNA with real-time fluorescent detection (RT-LAMP) with SYTO-82 dye on a model of herpes simplex virus (HSV) infection.
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