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.

Journal Articles

Antiviral effects of human placenta hydrolysate (Laennec) against SARS-CoV-2 in vitro and in the ferret model: Eun-Ha Kim , Young-il Kim , Seung-Gyu Jang , Minju Im , Kyeongsoo Jeong , Young Ki Choi , Hae-Jung Han; J. Microbiol. 2021;59(11):1056-1062. Published online October 6, 2021; DOI: https://doi.org/10.1007/s12275-021-1367-2

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The COVID-19 pandemic has caused unprecedented health, social, and economic crises worldwide. However, to date, there is an only a limited effective treatment for this disease. Human placenta hydrolysate (hPH) has previously been shown to be safe and to improve the health condition in patients with hyperferritinemia and COVID-19. In this study, we aimed to determine the antiviral effects of hPH against SARS-CoV-2 in vitro and in vivo models and compared with Remdesivir, an FDA-approved drug for COVID-19 treatment. To assess whether hPH inhibited SARS-CoV-2 replication, we determined the CC50, EC50, and selective index (SI) in Vero cells by infection with a SARS-CoV-2 at an MOI of 0.01. Further, groups of ferrets infected with 105.8 TCID50/ml of SARS-CoV-2 and treated with hPH at 2, 4, 6 dpi, and compared their clinical manifestation and virus titers in respiratory tracts with PBS control-treated group. The mRNA expression of immunerelated cytokines was determined by qRT-PCR. hPH treatment attenuated virus replication in a dose-dependent manner in vitro. In a ferret infection study, treatment with hPH resulted in minimal bodyweight loss and attenuated virus replication in the nasal wash, turbinates, and lungs of infected ferrets. In addition, qRT-PCR results revealed that the hPH treatment remarkably upregulated the gene expression of type I (IFN-α and IFN-β) and II (IFN-γ) IFNs in SARS-CoV-2 infected ferrets. Our data collectively suggest that hPH has antiviral efficacy against SARS-CoV-2 and might be a promising therapeutic agent for the treatment of SARS-CoV-2 infection.

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Development and validation of a novel RP-HPLC method for determination of Remdesivir: Investigation of the greenness for the proposed method
Karthika Paul, B.H. Jaswanth Gowda, R.S. Chandan
Results in Chemistry.2025; 16: 102382. CrossRef
Perinatal Hypoxia and Immune System Activation in Schizophrenia Pathogenesis: Critical Considerations During COVID-19 Pandemic
I Kawikova, K Hakenova, M Lebedeva, L Kleteckova, L Jakob, V Spicka, L Wen, F Spaniel, K Vales
Physiological Research.2024; : S615. CrossRef
Human Placenta Extract (HPH) Suppresses Inflammatory Responses in TNF-α/IFN-γ-Stimulated HaCaT Cells and a DNCB Atopic Dermatitis (AD)-Like Mouse Model
Jung Ok Lee, Youna Jang, A Yeon Park, Jung Min Lee, Kyeongsoo Jeong, So-Hyun Jeon, Hui Jin, Minju Im, Jae-Won Kim, Beom Joon Kim
Journal of Microbiology and Biotechnology.2024; 34(10): 1969. CrossRef
Systematic analysis of the pharmacology of standardized extracts of human placenta
T. E. Bogacheva, I. Yu. Torshin, O. A. Gromova
Pharmacokinetics and Pharmacodynamics.2024; (4): 3. CrossRef
Distinctive Combinations of RBD Mutations Contribute to Antibody Evasion in the Case of the SARS-CoV-2 Beta Variant
Tae-Hun Kim, Sojung Bae, Sunggeun Goo, Jinjong Myoung
Journal of Microbiology and Biotechnology.2023; 33(12): 1587. CrossRef
Current state-of-the-art and potential future therapeutic drugs against COVID-19
Ailong Sha, Yi Liu, Haiyan Hao
Frontiers in Cell and Developmental Biology.2023;[Epub] CrossRef
SARS-CoV-2 Aerosol and Intranasal Exposure Models in Ferrets
Elizabeth E. Zumbrun, Samantha E. Zak, Eric D. Lee, Philip A. Bowling, Sara I. Ruiz, Xiankun Zeng, Jeffrey W. Koehler, Korey L. Delp, Russel R. Bakken, Shannon S. Hentschel, Holly A. Bloomfield, Keersten M. Ricks, Tamara L. Clements, April M. Babka, John
Viruses.2023; 15(12): 2341. CrossRef
Human placenta hydrolysates: from V.P. Filatov to the present day: Review
Olga A. Gromova, Ivan Yu. Torshin, Alexander G. Chuchalin, Valeriy А. Maximov
Terapevticheskii arkhiv.2022; 94(3): 434. CrossRef

Differences in seroprevalence between epicenter and non-epicenter areas of the COVID-19 outbreak in South Korea: Hye Won Jeong , Hyun-Ha Chang , Eun Ji Kim , Yu Kyung Kim , Se-Mi Kim , Eun-Ha Kim , Young-Il Kim , Mark Anthony B. Casel , Seong-Gyu Kim , Rare Rollon , Seung-Gyu Jang , Kwang-Min Yu , Hee-Sung Kim , Hee Sue Park , Su-Jin Park , Yong-Dae Kim , Eung-Gook Kim , Young Ki Choi; J. Microbiol. 2021;59(5):530-533. Published online April 28, 2021; DOI: https://doi.org/10.1007/s12275-021-1095-7

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Abstract PDF

To compare the standardized severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) seroprevalence of high epicenter region with non-epicenter region, serological studies were performed with a total of 3,268 sera from Daegu City and 3,981 sera from Chungbuk Province. Indirect immunofluorescence assay (IFA) for SARS-CoV-2 IgG results showed a high seroprevalence rate in the Daegu City (epicenter) compared with a non-epicenter area (Chungbuk Province) (1.27% vs. 0.91%, P = 0.0358). It is noteworthy that the highest seroprevalence in Daegu City was found in elderly patients (70’s) whereas young adult patients (20’s) in Chungbuk Province showed the highest seroprevalence. Neutralizing antibody (NAb) titers were found in three samples from Daegu City (3/3, 268, 0.09%) while none of the samples from Chungbuk Province were NAb positive. These results demonstrated that even following the large outbreak, the seropositive rate of SARS-CoV-2 in the general population remained low in South Korea.

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Distinctive Combinations of RBD Mutations Contribute to Antibody Evasion in the Case of the SARS-CoV-2 Beta Variant
Tae-Hun Kim, Sojung Bae, Sunggeun Goo, Jinjong Myoung
Journal of Microbiology and Biotechnology.2023; 33(12): 1587. CrossRef
The Seroprevalence of SARS-CoV-2 in Children During Early COVID-19 Pandemic in Korea: A Nationwide, Population-Based Study
Jin Lee, Young June Choe, Dohsik Minn, Jong-Hyun Kim
Journal of Korean Medical Science.2022;[Epub] CrossRef

Development of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) thermal inactivation method with preservation of diagnostic sensitivity: Young-Il Kim , Mark Anthony B. Casel , Se-Mi Kim , Seong-Gyu Kim , Su-Jin Park , Eun-Ha Kim , Hye Won Jeong , Haryoung Poo , Young Ki Choi; J. Microbiol. 2020;58(10):886-891. Published online September 29, 2020; DOI: https://doi.org/10.1007/s12275-020-0335-6

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Abstract PDF

Various treatments and agents had been reported to inactivate RNA viruses. Of these, thermal inactivation is generally considered an effective and cheap method of sample preparation for downstream assays. The purpose of this study is to establish a safe inactivation method for SARS-CoV-2 without compromising the amount of amplifiable viral genome necessary for clinical diagnoses. In this study, we demonstrate the infectivity and genomic stability of SARSCoV- 2 by thermal inactivation at both 56°C and 65°C. The
results
substantiate that viable SARS-CoV-2 is readily inactivated when incubated at 56°C for 30 min or at 65°C for 10 min. qRT-PCR of specimens heat-inactivated at 56°C for 30 min or 65°C for 15 min revealed similar genomic RNA stability compared with non-heat inactivated specimens. Further, we demonstrate that 30 min of thermal inactivation at 56°C could inactivate viable viruses from clinical COVID-19 specimens without attenuating the qRT-PCR diagnostic sensitivity. Heat treatment of clinical specimens from COVID-19 patients at 56°C for 30 min or 65°C for 15 min could be a useful
method
for the inactivation of a highly contagious agent, SARS-CoV-2. Use of this method would reduce the potential for secondary infections in BSL2 conditions during diagnostic procedures. Importantly, infectious virus can be inactivated in clinical specimens without compromising the sensitivity of the diagnostic RT-PCR assay.

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Ming-Sian Wu, Pu-Chieh Chang, Po-Lin Lin, Chun-Hsi Tso, Hsin-Mei Chen, Yi-Hsuan Peng, Po-Chih Wu, Jia-Chuan Hsu, Der-Yuan Wang
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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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Abstract

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.

Citations

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Genetic Characterization of Kazakhstan Isolates: Avian Influenza H9N2 Viruses Demonstrate Their Potential to Infect Mammals
Barshagul Baikara, Kobey Karamendin, Yermukhammet Kassymbekov, Klara Daulbayeva, Temirlan Sabyrzhan, Sardor Nuralibekov, Yelizaveta Khan, Nurlan Sandybayev, Sasan Fereidouni, Aidyn Kydyrmanov
Viruses.2025; 17(5): 685. CrossRef
An Influenza A virus can evolve to use human ANP32E through altering polymerase dimerization
Carol M. Sheppard, Daniel H. Goldhill, Olivia C. Swann, Ecco Staller, Rebecca Penn, Olivia K. Platt, Ksenia Sukhova, Laury Baillon, Rebecca Frise, Thomas P. Peacock, Ervin Fodor, Wendy S. Barclay
Nature Communications.2023;[Epub] CrossRef
Current situation and control strategies of H9N2 avian influenza in South Korea
Mingeun Sagong, Kwang-Nyeong Lee, Eun-Kyoung Lee, Hyunmi Kang, Young Ki Choi, Youn-Jeong Lee
Journal of Veterinary Science.2023;[Epub] CrossRef
Antigenic Evolution Characteristics and Immunological Evaluation of H9N2 Avian Influenza Viruses from 1994–2019 in China
Qingzheng Liu, Lingcai Zhao, Yanna Guo, Yongzhen Zhao, Yingfei Li, Na Chen, Yuanlu Lu, Mengqi Yu, Lulu Deng, Jihui Ping
Viruses.2022; 14(4): 726. CrossRef
Molecular epidemiology and pathogenicity of H5N1 and H9N2 avian influenza viruses in clinically affected chickens on farms in Bangladesh
Ripatun Nahar Ripa, Joshua E. Sealy, Jayna Raghwani, Tridip Das, Himel Barua, Md. Masuduzzaman, A. K. M. Saifuddin, Md. Reajul Huq, Mohammad Inkeyas Uddin, Munir Iqbal, Ian Brown, Nicola S. Lewis, Dirk Pfeiffer, Guillaume Fournie, Paritosh Kumar Biswas
Emerging Microbes & Infections.2021; 10(1): 2223. CrossRef
Mouse adaptation of the H9N2 avian influenza virus causes the downregulation of genes related to innate immune responses and ubiquitin-mediated proteolysis in mice
Jing Guo, Xinxin Gao, Baotao Liu, Yubao Li, Wenqiang Liu, Jianbiao Lu, Cheng Liu, Rui Xue, Xuyong Li
Medical Microbiology and Immunology.2020; 209(2): 151. CrossRef
H9 Influenza Viruses: An Emerging Challenge
Silvia Carnaccini, Daniel R. Perez
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Adaptive amino acid substitutions enable transmission of an H9N2 avian influenza virus in guinea pigs
Liu Lina, Chen Saijuan, Wang Chengyu, Lu Yuefeng, Dong Shishan, Chen Ligong, Guo Kangkang, Guo Zhendong, Li Jiakai, Zhang Jianhui, Luo Qingping, Zhang Wenting, Shang Yu, Wang Honglin, Zhang Tengfei, Wen Guoyuan, Zhu Jiping, Zhang Chunmao, Jin Meilin, Gao
Scientific Reports.2019;[Epub] CrossRef
A PB1-K577E Mutation in H9N2 Influenza Virus Increases Polymerase Activity and Pathogenicity in Mice
Haruhiko Kamiki, Hiromichi Matsugo, Tomoya Kobayashi, Hiroho Ishida, Akiko Takenaka-Uema, Shin Murakami, Taisuke Horimoto
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Genetics and biological property analysis of Korea lineage of influenza A H9N2 viruses
Min Kang, Hyung-Kwan Jang
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The significance of avian influenza virus mouse-adaptation and its application in characterizing the efficacy of new vaccines and therapeutic agents
Won-Suk Choi, Khristine Kaith S. Lloren, Yun Hee Baek, Min-Suk Song
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Rapid acquisition of polymorphic virulence markers during adaptation of highly pathogenic avian influenza H5N8 virus in the mouse
Won-Suk Choi, Yun Hee Baek, Jin Jung Kwon, Ju Hwan Jeong, Su-Jin Park, Young-il Kim, Sun-Woo Yoon, Jungwon Hwang, Myung Hee Kim, Chul-Joong Kim, Richard J. Webby, Young Ki Choi, Min-Suk Song
Scientific Reports.2017;[Epub] CrossRef
Vaccine Efficacy of Inactivated, Chimeric Hemagglutinin H9/H5N2 Avian Influenza Virus and Its Suitability for the Marker Vaccine Strategy
Se Mi Kim, Young-Il Kim, Su-Jin Park, Eun-Ha Kim, Hyeok-il Kwon, Young-Jae Si, In-Won Lee, Min-Suk Song, Young Ki Choi, Jae U. Jung
Journal of Virology.2017;[Epub] CrossRef
Prevalence and diversity of H9N2 avian influenza in chickens of Northern Vietnam, 2014
Duong Mai Thuy, Thomas P. Peacock, Vu Thi Ngoc Bich, Thomas Fabrizio, Dang Nguyen Hoang, Nguyen Dang Tho, Nguyen Thi Diep, Minh Nguyen, Le Nguyen Minh Hoa, Hau Thi Thu Trang, Marc Choisy, Ken Inui, Scott Newman, Nguyen vu Trung, Rogier van Doorn, Thanh Lo
Infection, Genetics and Evolution.2016; 44: 530. CrossRef
PB2 subunit of avian influenza virus subtype H9N2: a pandemic risk factor
Hanna Sediri, Swantje Thiele, Folker Schwalm, Gülsah Gabriel, Hans-Dieter Klenk
Journal of General Virology.2016; 97(1): 39. CrossRef

Adjuvant Efficacy of mOMV against Avian Influenza Virus Infection in Mice: Byeong-Jae Lee , Sang-Ho Lee , Min-Suk Song , Philippe Noriel Q. Pascua , Hyeok-il Kwon , Su-Jin Park , Eun-Ha Kim , Arun Decano , Se Mi Kim , Gyo Jin Lim , Doo-Jin Kim , Kyu-Tae Chang , Sang-Hyun Kim , Young Ki Choi; J. Microbiol. 2013;51(5):682-688. Published online October 31, 2013; DOI: https://doi.org/10.1007/s12275-013-3411-3

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Abstract PDF: Highly pathogenic avian influenza H5N1 viruses are found chiefly in birds and have caused severe disease and death in infected humans. Development of influenza vaccines capable of inducing heterosubtypic immunity against a broad range of influenza viruses is the best option for the preparedness, since vaccination remains the principal method in controlling influenza viral infections. Here, a mOMV-adjuvanted recombinant H5N2 (rH5N2) whole virus antigen vaccine with A/Environment/Korea/W149/06(H5N1)-derived H5 HA and A/Chicken/Korea/ma116/04(H9N2)-derived N2 NA in the backbone of A/Puerto Rico/8/34(H1N1) was prepared and generated by reverse genetics. Groups of mice were vaccinated by a prime-boost regime with the rH5N2 vaccine (1.75 μg of HA with/without 10 μg mOMV or aluminum hydroxide adjuvant for comparison). At two weeks post-immunizations, vaccinated mice were challenged with lethal doses of 103.5 EID50/ml of H5N1 or H9N2 avian influenza viruses, and were monitored for 15 days. Both mOMV- and alum-adjuvant vaccine groups had high survival rates after H5N1 infection and low levels of body weight changes compared to control groups. Interestingly, the mOMV-adjuvanted group induced better cross-reactive antibody responses serologically and promoted cross-protectivity against H5N1 and H9N2 virus challenges. Our results suggest that mOMV could be used as a vaccine adjuvant in the development of effective vaccines used to control influenza A virus transmission.

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