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17 results on '"Shu, Meng-Hooi"'

7. Early detection of the dengue virus using reverse transcription-recombinase polymerase amplification

Author

Teoh, Boon-Teong, Sam, Sing-Sin, Tan, Kim-Kee, Danlami, Mohammed Bashar, Shu, Meng-Hooi, Johari, Jefree, Hooi, Poh-Sim, Brooks, David, Piepenburg, Olaf, Nentwich, Oliver, Wilder-Smith, Annelies, Franco, Leticia, Tenorio, Antonio, and AbuBakar, Sazaly

Subjects
Folkhälsovetenskap, global hälsa, socialmedicin och epidemiologi, Public Health, Global Health, Social Medicine and Epidemiology
Abstract

A method for the rapid diagnosis of early dengue virus (DENV) infection is highly needed. Here, a prototype reverse transcription-recombinase polymerase amplification (RT-RPA) assay was developed. The assay detected DENV RNA in Accepted manuscript posted online 7 January 2015, Free via Open Access: OA. The authors have paid a fee to allow immediate free access to this article.

Published
2015

11. Early Detection of Dengue Virus by Use of Reverse Transcription-Recombinase Polymerase Amplification

Author

Teoh, Boon-Teong, primary, Sam, Sing-Sin, additional, Tan, Kim-Kee, additional, Danlami, Mohammed Bashar, additional, Shu, Meng-Hooi, additional, Johari, Jefree, additional, Hooi, Poh-Sim, additional, Brooks, David, additional, Piepenburg, Olaf, additional, Nentwich, Oliver, additional, Wilder-Smith, Annelies, additional, Franco, Leticia, additional, Tenorio, Antonio, additional, and AbuBakar, Sazaly, additional

Published
2015
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16. Insertion of single-chain variable fragment (scFv) peptide linker improves surface display of influenza hemagglutinin (HA1) on non-recombinant Lactococcus lactis.

Author

Jee, Pui‐Fong, Chen, Fez‐Shin, Shu, Meng‐Hooi, Wong, Won Fen, Abdul Rahim, Raha, AbuBakar, Sazaly, and Chang, Li‐Yen

Subjects
INFLUENZA viruses, PEPTIDES, N-Acetylmuramidase, BIOTECHNOLOGY, HEMAGGLUTININ
Abstract

Heterologous protein displayed on the surface of Lactococcus lactis using the binding domain of N-acetylmuramidase (AcmA) has a potential application in vaccine delivery. In this study, we developed a non-recombinant L. lactis surface displaying the influenza A (H1N1) 2009 hemagglutinin (HA1). Three recombinant proteins, HA1/L/AcmA, HA1/AcmA, and HA1 were overexpressed in Escherichia coli, and purified. In the binding study using flow cytometry, the HA1/L/AcmA, which contained the single-chain variable fragment (scFv) peptide linker showed significantly higher percentage of binding counts and mean fluorescence binding intensity (MFI) (51.7 ± 1.4% and 3,594.0 ± 675.9, respectively) in comparison to the HA1/AcmA without the scFv peptide linker (41.1 ± 1.5% and 1,652.0 ± 34.1, respectively). Higher amount of HA1/L/AcmA (∼2.9 × 104 molecules per cell) was displayed on L. lactis when compared to HA1/AcmA (∼1.1 × 104 molecules per cell) in the immunoblotting analysis. The HA1/L/AcmA completely agglutinated RBCs at comparable amount of protein to that of HA1/AcmA and HA1. Computational modeling of protein structures suggested that scFv peptide linker in HA1/L/AcmA kept the HA1 and the AcmA domain separated at a much longer distance in comparison to HA1/AcmA. These findings suggest that insertion of the scFv peptide linker between HA1 and AcmA improved binding of recombinant proteins to L. lactis. Hence, insertion of scFv peptide linker can be further investigated as a potential approach for improvement of heterologous proteins displayed on the surface of L. lactis using the AcmA binding domain. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:154-162, 2017 [ABSTRACT FROM AUTHOR]

Published
2017
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17. Early Detection of Dengue Virus by Use of Reverse Transcription-Recombinase Polymerase Amplification

Author

Teoh, Boon-Teong, Sam, Sing-Sin, Tan, Kim-Kee, Danlami, Mohammed Bashar, Shu, Meng-Hooi, Johari, Jefree, Hooi, Poh-Sim, Brooks, David, Piepenburg, Olaf, Nentwich, Oliver, Wilder-Smith, Annelies, Franco, Leticia, Tenorio, Antonio, and AbuBakar, Sazaly

Abstract

ABSTRACTA method for the rapid diagnosis of early dengue virus (DENV) infection is highly needed. Here, a prototype reverse transcription-recombinase polymerase amplification (RT-RPA) assay was developed. The assay detected DENV RNA in <20 min without the need for thermocycling amplification. The assay enabled the detection of as few as 10 copies of DENV RNA. The designed RT-RPA primers and exoprobe detected the DENV genome of at least 12 genotypes of DENV circulating globally without cross-reacting with other arboviruses. We assessed the diagnostic performance of the RT-RPA assay for the detection of DENV RNA in 203 serum samples of patients with clinically suspected dengue. The sera were simultaneously tested for DENV using a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay, quantitative RT-PCR (qRT-PCR), and IgM- and IgG-capture enzyme-linked immunosorbent assays (ELISA). Acute DENV infection was confirmed in 130 samples and 61 of the samples (46.9%) were classified as viremic with qRT-PCR. The RT-RPA assay showed good concordance (? of =0.723) with the RT-LAMP and qRT-PCR assays in detecting the dengue viremic samples. When used in combination with ELISA, both the RT-RPA and RT-LAMP assays increased the detection of acute DENV infection to =95.7% (=45/47) in samples obtained within 5 days of illness. The results from the study suggest that the RT-RPA assay is the most rapid molecular diagnostic tool available for the detection of DENV. Hence, it is possible to use the RT-RPA assay in a laboratory to complement routine serology testing for dengue.

Published
2014
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