Journal of Microbiology

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Development of an RT-LAMP−CRISPR/Cas12a assay for rapid and specific detection of Bandavirus dabieense: Bo Seung Song, Yun Hee Baek, Eun-Ha Kim, Hyeok-Il Kwon, Ah-Hyeon Kim, Si-Hyun Lee, Yu-Bin Son, Soo-Hyeon Kim, Min-Suk Song, Young Ki Choi, Su-Jin Park; J. Microbiol. 2025;63(11):e2506013. Published online November 30, 2025; DOI: https://doi.org/10.71150/jm.2506013

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Abstract PDF: Bandavirus dabieense, a single-stranded RNA virus, is the causative agent of severe fever with thrombocytopenia syndrome (SFTS), a disease associated with high fatality rates. Early and accurate diagnosis is essential for improving clinical outcomes, particularly given the limited therapeutic options and high mortality rates associated with SFTS. However, while highly sensitive, conventional diagnostic methods such as PCR and qRT-PCR require specialized laboratory facilities and trained personnel, making them impractical for rapid detection in resource-limited settings. To address these challenges, we developed a rapid and highly sensitive assay for Bandavirus dabieense detection by integrating reverse transcription loop-mediated isothermal amplification (RT-LAMP) with CRISPR/Cas12a technology. LAMP primers and guide RNA sequences were designed to target the L gene, ensuring broad detection across viral genotypes. The optimized assay demonstrated a detection limit of 5 RNA copies per reaction, showing more sensitivity than qRT-PCR, and exhibited 100% concordance with qRT-PCR results in clinical samples. Given its speed, accuracy, and field applicability, this LAMP-CRISPR/Cas12a-based assay represents a promising diagnostic tool for early SFTSV detection, particularly in resource-constrained environments where conventional molecular diagnostics are not readily available.

Research Support, Non-U.S. Gov'ts

Molecular characterization of mammalian-adapted Korean-type avian H9N2 virus and evaluation of its virulence in mice: Kuk Jin Park , Min-Suk Song , Eun-Ha Kim , Hyeok-il Kwon , Yun Hee Baek , Eun-hye Choi , Su-Jin Park , Se Mi Kim , Young-il Kim , Won-Suk Choi , Dae-Won Yoo , Chul-Joong Kim , Young Ki Choi; J. Microbiol. 2015;53(8):570-577. Published online July 31, 2015; DOI: https://doi.org/10.1007/s12275-015-5329-4

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Avian influenza A virus (AIV) is commonly isolated from domestic poultry and wild migratory birds, and the H9N2 subtype is the most prevalent and the major cause of severe disease in poultry in Korea. In addition to the veterinary concerns regarding the H9N2 subtype, it is also considered to be the next potential human pandemic strain due to its rapid evolution and interspecies transmission. In this study, we utilize serial lung-to-lung passage of a low pathogenic avian influenza virus (LPAI) H9N2 (A/Ck/Korea/163/04, WT163) (Y439-lineage) in mice to increase pathogenicity and investigate the potential virulence marker. Mouse-adapted H9N2 virus obtained high virulence (100% mortality) in mice after 98 serial passages. Sequence results show that the mouse adaptation (ma163) possesses several mutations within seven gene segments (PB2, PA, HA, NP, NA, M, and NS) relative to the wild-type strain. The HA gene showed the most mutations (at least 11) with one resulting in the loss of an N-glycosylation site (at amino acid 166). Moreover, reverse genetic studies established that an E627K substitution in PB2 and the loss of the N-glycosylation site in the HA protein (aa166) are critical virulence markers in the mouse-adapted H9N2 virus. Thus, these results add to the increasing body of mutational analysis data defining the function of the viral polymerase and HA genes and their roles in mammalian host adaptation. To our knowledge, this is first report of the generation of a mammalian-adapted Korea H9N2 virus (Y493-lineages). Therefore, this study offers valuable insights into the molecular evolution of the LPAI Korean H9N2 in a new host and adds to the current knowledge of the molecular markers associated with increased virulence.

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Molecular Characterization and Phylogenetic Analysis of H3N2 Human Influenza A Viruses in Cheongju, South Korea: Yun Hee Baek , Jeung Hyun Park , Young Jun Song , Min-Suk Song , Philippe Noriel Q. Pascua , Yoon-Soo Hahn , Heon-Seok Han , Ok-Jun Lee , Ki-Soon Kim , Chun Kang , Young-Ki Choi; J. Microbiol. 2009;47(1):91-100. Published online February 20, 2009; DOI: https://doi.org/10.1007/s12275-008-0207-y

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Abstract PDF

To investigate the genetic characteristics of human influenza viruses circulating in Chungbuk province, we tested 510 clinical samples of nasopharyngeal suction from pediatric patients diagnosed with respiratory illness between June 2007 and June 2008. Genetic characterization of the HA genes of H3N2 isolates indicated the relative higher similarity to A/Virginia/04/07 (99.6%) rather than that of A/Wisconsin/67/2005 (98.4%), a Northern Hemisphere 2007~2008 vaccine strain, based on amino acid sequences. We found several altered amino acids at the H3 HA1 antigenic sites compared with the vaccine strain; K140I at site A, K158R at site B, and K173N (H471) or K173Q, and S262N at site E, but there was no antigenic shift among the H3N2 viruses. Interestingly, A/Cheongju/H383/08 and A/Cheongju/H407/08 isolates had single amino acid substitution at D151G on the catalytic site of the N2 NA while A/Cheongju/H412/08 and A/Cheongju/H398/07 isolates had one amino acid deletion at residue 146. Furthermore, we found that 25% (3 out of 12 isolates) of the H3N2 subtype viruses had the amino acid substitution at position 31 on the M2 protein (Aspartic acid to Asparagine) and confirmed their drug-resistance by biological assays. Taken together, the results of this study demonstrated continuous evolutions of human H3N2 viruses by antigenic drift and also highlighted the need to closely monitor antigenic drug resistance in influenza A viruses to aid in the early detection of potentially pandemic strains, as well as underscore the need for new therapeutics.

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Evaluation of the Efficacy of a Pre-pandemic H5N1 Vaccine (MG1109) in Mouse and Ferret Models: Min-Suk Song , Ho-Jin Moon , Hyeok-il Kwon , Philippe Noriel Q. Pascua , Jun Han Lee , Yun Hee Baek , Kyu-Jin Woo , Juhee Choi , Sangho Lee , Hyunseung Yoo , In gyeong Oh , Yeup Yoon , Jong-Bok Rho , Moon-Hee Sung , Seung-Pyo Hong , Chul-Joong Kim , Young Ki Choi; J. Microbiol. 2012;50(3):487-488.

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Abstract PDF: The threat of a highly pathogenic avian influenza (HPAI) H5N1 virus causing the next pandemic remains a major concern. In this study, we evaluated the immunogenicity and efficacy of an inactivated whole-virus H5N1 pre-pandemic vaccine (MG1109) formulated by Green Cross Co., Ltd containing the hemagglutinin (HA) and neuraminidase (NA) genes of the clade 1 A/Vietnam/1194/04 virus in the backbone of A/Puerto Rico/8/34 (RgVietNam/04xPR8/34). Administration of the MG1109 vaccine (2-doses) in mice and ferrets elicited high HI and SN titers in a dose-dependent manner against the homologous (RgVietNam/04xPR8/34) and various heterologous H5N1 strains, (RgKor/W149/06xPR8/34, RgCambodia/04xPR8/34, RgGuangxi/05xPR8/34), including a heterosubtypic H5N2 (A/Aquatic bird/orea/W81/05) virus. However, efficient cross-reactivity was not observed against heterosubtypic H9N2 (A/Ck/Korea/H0802/08) and H1N1 (PR/8/34) viruses. Mice immunized with 1.9 μg HA/dose of MG1109 were completely protected from lethal challenge with heterologous wild-type HPAI H5N1 A/EM/Korea/W149/06 (clade 2.2) and mouse-adapted H5N2 viruses. Furthermore, ferrets administered at least 3.8 μg HA/dose efficiently suppressed virus growth in the upper respiratory tract and lungs. Vaccinated mice and ferrets also demonstrated attenuation of clinical disease signs and limited virus spread to other organs. Thus, this vaccine provided immunogenic responses in mouse and ferret models even against challenge with heterologous HPAI H5N1 and H5N2 viruses. Since the specific strain of HPAI H5N1 virus that would potentially cause the next outbreak is unknown, pre-pandemic vaccine preparation that could provide crossprotection against various H5 strains could be a useful approach in the selection of promising candidate vaccines in the future.

ERRATUM] Evaluation of the Efficacy of a Pre-pandemic H5N1 Vaccine (MG1109) in Mouse and Ferret Models: Min-Suk Song , Ho-Jin Moon , Hyeok-il Kwon , Philippe Noriel Q. Pascua , Jun Han Lee , Yun Hee Baek , Kyu-Jin Woo , Juhee Choi , Sangho Lee , Hyunseung Yoo , In gyeong Oh , Yeup Yoon , Jong-Bok Rho , Moon-Hee Sung , Seung-Pyo Hong , Chul-Joong Kim , Young Ki Choi; J. Microbiol. 2012;50(4):715-715.

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Abstract PDF: In the article by Song et al. that appears in the Journal of Microbiology 2012; 50, 478-488. Page 478, the name of 7th author, Kyu-Jin Woo, should read as Gyu-Jin Woo.

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