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Start Over Author "Hisanori Inoue" Topic cytomegalovirus
2 results on '"Hisanori Inoue"'

1. Multiplex PCR-based DNA array for simultaneous detection of three human herpesviruses, EVB, CMV and KSHV.

Author

Fujimuro M, Nakaso K, Nakashima K, Sadanari H, Hisanori I, Teishikata Y, Hayward SD, and Yokosawa H

Subjects
DNA Probes genetics, DNA, Viral genetics, Fibroblasts virology, Genome, Viral genetics, Humans, Nucleic Acid Hybridization, Oligonucleotide Array Sequence Analysis, Polymerase Chain Reaction, Sensitivity and Specificity, Cytomegalovirus genetics, Cytomegalovirus isolation & purification, DNA, Viral analysis, Herpesvirus 4, Human genetics, Herpesvirus 4, Human isolation & purification, Herpesvirus 8, Human genetics, Herpesvirus 8, Human isolation & purification
Abstract

Human lymphotropic herpesviruses, Epstein-Barr virus (EBV), cytomegalovirus (CMV) and Kaposi's sarcoma-associated herpesvirus (KSHV) are responsible for a wide variety of human diseases. Due to an increase in diseased states associated with immunosuppression, more instances of co-morbid infections with these herpesviruses have resulted in viral reactivations that have caused numerous fatalities. Therefore, the development of rapid and accurate method to detect these viruses in immunocompromised patients is vital for immediate treatment with antiviral prophylactic drugs. In this study, we developed a new multiplex PCR method coupled to DNA array hybridization, which can simultaneously detect all three human herpesviruses in one single cell sample. Multiplex PCR primers were designed to amplify specific regions of the EBV (EBER1), CMV (IE) and KSHV (LANA) viral genomes. Pre-clinical application of this method revealed that this approach is capable of detecting as few as 1 copy of the viral genomes for KSHV and CMV and 100 copies of the genome for EBV. Furthermore, this highly sensitive test showed no cross-reactivity among the three viruses and is capable of detecting both KSHV and EBV viral genomes simultaneously in the lymphoblastoid cells that have been double infected with both viruses. Thus, this array-based approach serves as a rapid and reliable diagnostic tool for clinical applications.

Published
2006
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2. Epidemiology of Epstein–Barr virus, cytomegalovirus, and kaposi's sarcoma-associated herpesvirus infections in peripheral blood leukocytes revealed by a multiplex PCR assay

Author

Kazuhiro Nakaso, Hideyoshi Yokosawa, Hitoshi Sasajima, Tomohiro Yamamoto, Masahiro Fujimuro, Morie Nishiwaki, Yoshie Fujiwara, Kenji Nakashima, Yasuhiro Teishikata, Hidetaka Sadanari, Hisanori Inoue, and Naoki Ogawa

Subjects
Epstein-Barr Virus Infections, Herpesvirus 4, Human, viruses, Cytomegalovirus, Blood Donors, Biology, medicine.disease_cause, Polymerase Chain Reaction, Sensitivity and Specificity, Virus, Cell Line, Plasmid, hemic and lymphatic diseases, Virology, Multiplex polymerase chain reaction, Leukocytes, Prevalence, medicine, Kaposi's sarcoma-associated herpesvirus, Sarcoma, Kaposi, virus diseases, Herpesviridae Infections, biochemical phenomena, metabolism, and nutrition, medicine.disease, Molecular biology, Epstein–Barr virus, Lymphoma, Infectious Diseases, Cytomegalovirus Infections, DNA, Viral, Herpesvirus 8, Human, Primary effusion lymphoma
Abstract

A multiplex polymerase chain reaction (PCR) has been developed for the simultaneous detection of Epstein-Barr virus (EBV), cytomegalovirus (CMV), and Kaposi's sarcoma-associated herpesvirus (KSHV) in a clinical sample. Primers of multiplex PCR were designed to amplify specific regions of the EBV EBNA1, CMV IE2, and KSHV LANA genes. This multiplex PCR assay was found to have detection sensitivities of 1-10 copies of purified viral DNA cloned into the plasmid. To assess diagnostic and pre-clinical applications with this method, we utilized KSHV-positive primary effusion lymphoma (PEL) cells, EBV-positive Burkitt's lymphoma cells, CMV-infected fibroblast cells, and clinically prepared peripheral blood leukocytes (PBLs) that had been infected with viruses. We found that this multiplex PCR assay has high sensitivity and specificity for simultaneous detection of EBV, CMV, and KSHV genomes in a single amplification from a clinical material. Using this multiplex PCR assay, we investigated the prevalence of EBV, CMV, and KSHV in PBL samples from normal Japanese randomly selected. KSHV, EBV, and CMV genomes were detected in samples from 2 (0.2%), 377 (39.5%), and 27 (2.8%) of the 953 blood donors, respectively. Interestingly, both EBV and CMV genomes were detected in samples from all KSHV-positive donors.

Published
2006

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