Visual detection and evaluation of latent and lytic gene expression during Epstein-Barr virus infection using one-step reverse transcription loop-mediated isothermal amplification

International Journal of Molecular Sciences
Xiaoying LiuYefu Wang

Abstract

Epstein-Barr virus (EBV)-associated disease exhibits distinct gene expression patterns characterized by the transcription of EBV nuclear antigen (EBNA) 1, EBNA2, latent membrane protein (LMP) 1, LMP2A, and BZLF1 (Zebra). A series of visual reverse transcript loop-mediated isothermal amplification (RT-LAMP) assays were performed to examine the expression of EBNA1, EBNA2, LMP1, LMP2A and BZLF1. The sensitivity of RT-LAMP for these transcripts was approximately equivalent to real-time RT-PCR (RT-qPCR), which was developed to quantify relative levels of EBV transcripts, and 10 to 100-fold more sensitive than conventional RT-PCR. Cross-reactions to other viruses were not observed upon examination of cell lines infected with herpes simplex viruses-1 and -2 (HSV-1 and -2), varicella zoster virus (VZV), human cytomegalovirus (HCMV) or Kaposi's sarcoma-associated herpesvirus. When applied to 146 specimens, RT-LAMP exhibited high clinical sensitivity and specificity, with an excellent agreement (κ > 0.92) compared to RT-qPCR. These assays are convenient for rapid early diagnosis and for surveillance of EBV-infected individuals by evaluating the EBV transcriptional profile, because the results can be visualized with the naked eye. These a...Continue Reading

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Citations

Aug 20, 2019·Indian Journal of Medical Microbiology·Mini P SinghR K Ratho
Jan 1, 2019·Indian Journal of Medical Microbiology·Mini P SinghR K Ratho

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Datasets Mentioned

BETA
M12553.1
K03333.1
AF023171.1
GU979730.1
and
EU340368.1
BC023632.2
GU979791.1
the
AY825078.1

Methods Mentioned

BETA
PCR
electrophoresis
fluorescence assay
ELISA

Software Mentioned

Primer Explorer
SPSS

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