tetano
Editor, Senior Moderator
J Virol Methods. 2019 Dec 12:113801. doi: 10.1016/j.jviromet.2019.113801. [Epub ahead of print] [h=1]A Clinic-Based Direct Real-time Fluorescent Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Influenza Virus.[/h]
Ngan LT[SUP]1[/SUP], Takayama I[SUP]2[/SUP], Binh NG[SUP]1[/SUP], Phuong TT[SUP]1[/SUP], Tuong Van VT[SUP]1[/SUP], Vuong BM[SUP]1[/SUP], Co DX[SUP]1[/SUP], Dung LT[SUP]1[/SUP], Phuong PT[SUP]1[/SUP], Cuong DD[SUP]1[/SUP], Thach PT[SUP]1[/SUP], Van Thanh D[SUP]1[/SUP], Phuong Thuy PT[SUP]3[/SUP], Chau NQ[SUP]1[/SUP], Tuan DQ[SUP]1[/SUP], Takasaki J[SUP]4[/SUP], Semba S[SUP]5[/SUP], Odagiri T[SUP]2[/SUP], Nakajima N[SUP]6[/SUP], Kageyama T[SUP]2[/SUP].
[h=3]Author information[/h] 1 Bach Mai Hospital, 78 Giai Phong Street, Dong Da District, Hanoi, Viet Nam. 2 Influenza Virus Research Center, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama-shi, Tokyo, 208-0011, Japan. 3 NCGM-Bach Mai Hospital Medical Collaboration Center, 78 Giai Phong Street, Dong Da District, Hanoi, Viet Nam. 4 National Center for Global Health and Medicine, 1-21-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan. 5 Eiken Chemical Co., Ltd, 4-19-9 Taito, Taito-ku, Tokyo, 110-8408, Japan. 6 Department of Pathology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan. Electronic address: tenko@nih.go.jp.
[h=3]Abstract[/h] Seasonal influenza virus causes acute respiratory tract infections, which can be severe in children and the elderly. At present, rapid influenza diagnostic tests (RIDTs) are popular at clinical sites because they enable early diagnosis and avoid unnecessary use of antibiotics; in addition, high risk patients with underlying disease can be given antiviral drugs. However, the sensitivity and specificity of some of those tests are relatively poor. To overcome these problems, nucleic acid-based molecular point-of-care tests have been developed; however, they are significantly more expensive than RIDTs. Previously, the authors developed real-time reverse transcription loop-mediated isothermal amplification (rRT-LAMP) assays using a quenching primer to detect influenza viruses. However, the assay is limited to laboratory use because it requires a nucleic acid purification step and preparation of reaction mixtures on ice. Therefore, the authors developed and validated direct rRT-LAMP assays that require no nucleic acid purification steps using commercial RNA isolation kits, and no storage and handling of reagents on ice. These assays can be performed within 10-30 min and require only mixing a clinical specimen with extraction reagent followed by addition of a lyophilized detection reagent. The established assay showed high sensitivity and specificity when validated using 310 clinical specimens. Thus, the assay is a powerful tool for molecular diagnosis of seasonal influenza virus infection in the clinic.
Copyright ? 2019. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] RT-LAMP assay; clinical site; influenza virus; rapid; real-time
PMID: 31838003 DOI: 10.1016/j.jviromet.2019.113801
Ngan LT[SUP]1[/SUP], Takayama I[SUP]2[/SUP], Binh NG[SUP]1[/SUP], Phuong TT[SUP]1[/SUP], Tuong Van VT[SUP]1[/SUP], Vuong BM[SUP]1[/SUP], Co DX[SUP]1[/SUP], Dung LT[SUP]1[/SUP], Phuong PT[SUP]1[/SUP], Cuong DD[SUP]1[/SUP], Thach PT[SUP]1[/SUP], Van Thanh D[SUP]1[/SUP], Phuong Thuy PT[SUP]3[/SUP], Chau NQ[SUP]1[/SUP], Tuan DQ[SUP]1[/SUP], Takasaki J[SUP]4[/SUP], Semba S[SUP]5[/SUP], Odagiri T[SUP]2[/SUP], Nakajima N[SUP]6[/SUP], Kageyama T[SUP]2[/SUP].
[h=3]Author information[/h] 1 Bach Mai Hospital, 78 Giai Phong Street, Dong Da District, Hanoi, Viet Nam. 2 Influenza Virus Research Center, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama-shi, Tokyo, 208-0011, Japan. 3 NCGM-Bach Mai Hospital Medical Collaboration Center, 78 Giai Phong Street, Dong Da District, Hanoi, Viet Nam. 4 National Center for Global Health and Medicine, 1-21-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan. 5 Eiken Chemical Co., Ltd, 4-19-9 Taito, Taito-ku, Tokyo, 110-8408, Japan. 6 Department of Pathology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan. Electronic address: tenko@nih.go.jp.
[h=3]Abstract[/h] Seasonal influenza virus causes acute respiratory tract infections, which can be severe in children and the elderly. At present, rapid influenza diagnostic tests (RIDTs) are popular at clinical sites because they enable early diagnosis and avoid unnecessary use of antibiotics; in addition, high risk patients with underlying disease can be given antiviral drugs. However, the sensitivity and specificity of some of those tests are relatively poor. To overcome these problems, nucleic acid-based molecular point-of-care tests have been developed; however, they are significantly more expensive than RIDTs. Previously, the authors developed real-time reverse transcription loop-mediated isothermal amplification (rRT-LAMP) assays using a quenching primer to detect influenza viruses. However, the assay is limited to laboratory use because it requires a nucleic acid purification step and preparation of reaction mixtures on ice. Therefore, the authors developed and validated direct rRT-LAMP assays that require no nucleic acid purification steps using commercial RNA isolation kits, and no storage and handling of reagents on ice. These assays can be performed within 10-30 min and require only mixing a clinical specimen with extraction reagent followed by addition of a lyophilized detection reagent. The established assay showed high sensitivity and specificity when validated using 310 clinical specimens. Thus, the assay is a powerful tool for molecular diagnosis of seasonal influenza virus infection in the clinic.
Copyright ? 2019. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] RT-LAMP assay; clinical site; influenza virus; rapid; real-time
PMID: 31838003 DOI: 10.1016/j.jviromet.2019.113801