論文 - 岡林 環樹
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Development of a direct duplex real-time PCR assay for rapid detection of domestic cat hepadnaviru 査読あり
Maya Shofa 1 2 , Akiho Ohkawa, Tamaki Okabayashi , Yasuyuki Kaneko , Akatsuki Saito
J.Vet. Diagn. Invest. 2023年2月
掲載種別：研究論文（学術雑誌）
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Development of a direct duplex real-time PCR assay for rapid detection of domestic cat hepadnavirus 査読あり
Maya Shofa 1 2 , Akiho Ohkawa 1 , Tamaki Okabayashi 1 2 3 , Yasuyuki Kaneko 4 , Akatsuki Saito 1 2 3
I. Vet. Diagn. Invest. 2023年2月
掲載種別：研究論文（学術雑誌）
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Mekata Hirohisa, Kawaguchi Takeshi, Iwao Kosho, Umeki Kazumi, Yamada Kentaro, Umekita Kunihiko, Okabayashi Tamaki
Japanese Journal of Medical Science and Biology advpub ( 0 ) 211 - 214 2023年1月
担当区分：責任著者 記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：国立感染症研究所
Severe fever with thrombocytopenia syndrome virus (SFTSV) is a causative agent of SFTS. Although SFTS is a fatal tick-borne zoonosis, the virus can infect humans without tick bite exposure. Recently, direct SFTSV transmission from companion pets to humans has become a significant problem. Herein, we reported a case of SFTSV transmission from a dead community cat to a woman who buried the cat in Miyazaki prefecture, Japan. The community cat died without a diagnosis of SFTS, and the woman buried it without precaution. Nine days later, the woman developed symptoms of SFTS. SFTS viral RNA and anti-SFTSV antibodies were detected in the woman. The animal carcass was exhumed, and tissue samples were collected to confirm viral infection in the cat. High copy numbers of viral RNA were detected. Sequences of the SFTSV M segment in samples from the cat and the woman showed 100% homology. The woman stated that she had touched blood that had leaked from the body when she buried it. On the other hand, the woman could have been infected when she took the cat to the animal hospital. This study highlights the risk of SFTSV infection via contact with sick or dead community cats.
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特集 重症熱性血小板減少症候群(SFTS)研究における最新知見--感染経路,病態,予防,治療 自然界におけるSFTSウイルス伝播様式
岡林 環樹
医学のあゆみ 284 ( 2 ) 129 - 132 2023年1月
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Giangaspero M., Okabayashi T.
Open Veterinary Journal 13 ( 4 ) 400 - 406 2023年
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Open Veterinary Journal
Background: In 2015, an unprecedented epidemic of microcephaly occurred in Brazil. Preliminary observations suggested the involvement of cofactors in the etiopathology of Zika virus-associated microcephaly. Bovine viral diarrhea virus (BVDV) was identified in fetal samples with microcephaly, originating in the state of Paraíba, and two virus sequences, obtained from the amniotic fluid collected from mothers with babies affected by Zika and microcephaly, have been characterized as two different species of BVDV, types 1 and 2. Aim: The involvement of BVDV as a co-factor in the etiopathogenesis of Zika virus-associated microcephaly was explored. Methods: A serological screening using an ELISA test was undertaken to detect antibodies against BVDV among patients referred to the Central Laboratory of Natal, Rio Grande do Norte, encompassing microcephalic babies and their mothers, mothers and pregnants not associated with microcephaly and general patients as a control group. Results: Two samples were positive out of 382 tested (0.52%). No specific relation with birth defects could be established. Conclusions: The study might suggest serological evidence of BVDV in humans. Further studies and the application of improved diagnostic tests adapted to humans are necessary to clarify the epidemiological extent and impact of BVDV.
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Wiradana P.A., Maharani A.Y., Sani M.D., Susilo R.J.K., Riandi M.I., Widhiantara I.G., Sudaryatma P.E., Okabayashi T., Mukti A.T.
IOP Conference Series: Earth and Environmental Science 1273 ( 1 ) 2023年
掲載種別：研究論文（学術雑誌） 出版者・発行元：IOP Conference Series: Earth and Environmental Science
The aim of the research is to monitor the dynamics of microbial number, water quality, and production performance in Pacific white shrimp (Litopenaeus vannamei) cultivation in Bulukumba, South Sulawesi Province, Indonesia, utilizing an intensive system. This research was carried out on seven Pacific white shrimp ponds over the duration of one maintenance cycle (84 days), from November 2019 to February 2020. Interestingly, the results showed that the highest total bacterial counts (TBC) were obtained in pond P3, namely in the range of 0.12-98.0×105 CFU/mL, the highest total Vibrio counts (TVC) were obtained in pond P5, namely in the range of 0.09-2.8×105 CFU/mL, luminous bacterial counts (LBC) were only found in ponds P1, P4, and P5 with the same range values, namely 0-100×103 CFU. The best performance of Pacific white shrimp production was found in pond P2, with a percentage of survival rate (SR) and productivity of 65 percent and 4074.5kg/ha, respectively, and the worst was found in pond P5, with a percentage of survival rate (SR) and productivity of 35 percent and 2583.66 kg/ha, respectively. Finally, the intense culture system used for Pacific white shrimp accumulates dynamics that may alter microbiology, water quality, and production performance.
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Inagaki H., Goto Y., Sugiyama H., Saito A., Okabayashi T., Watanabe K., Fujimoto S.
Frontiers in Public Health 10 1053729 2022年12月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Frontiers in Public Health
Reliable disinfection and sterilization technologies are needed to deal with the various infectious diseases spreading around the world. Furthermore, bacteria that are difficult to eliminate by ordinary disinfection are also a problem in the medical environment. We examined the germicidal effect of a newly developed deep-ultraviolet light-emitting diode (DUV-LED) prototype device (wavelength of 280 ± 5 nm; power of 0.9 to 1.4 mW/cm2) for floor sterilization against Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), Mycobacterium gordonae (M. gordonae), and Bacillus subtilis (B. subtilis). This prototype device is equipped with highly practical DUV-LEDs with a high output efficiency and a long life, and was designed with consideration of the irradiation distance and the angle of the DUV-LEDs to provide a uniform irradiation rate. We found a statistically significant reduction of ≥90% in the infectious titers of both E. coli and S. aureus after irradiation for 2 s. Although acid-fast bacilli and spore-type bacilli are generally thought to be resistant to UV light irradiation compared to general bacteria, the acid-fast bacillus M. gordonae was inactivated after irradiation for 10 s, and spore-type cells of the bacillus B. subtilis were inactivated by ≥90% after irradiation for 30 s. We also found that the effects were cumulative when irradiation was performed at intervals. In the future, the usefulness of this device as an infection control measure will be evaluated in daily medical practice.
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Generation of a bovine cell line for gene engineering using an HIV-1-based lentiviral vector
Morizako N., Butlertanaka E.P., Tanaka Y.L., Shibata H., Okabayashi T., Mekata H., Saito A.
Scientific Reports 12 ( 1 ) 16952 2022年12月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Scientific Reports
Human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors are indispensable tools for gene engineering in mammalian cells. Conversely, lentiviral vector transduction is severely inhibited in bovine cells. Previous studies demonstrated that this inhibition is caused by the anti-lentiviral host factor tripartite motif containing 5 (TRIM5), which targets incoming HIV-1 virions by interacting with the viral capsid. In this study, we investigated several methods for overcoming the limited applicability of lentiviral vectors in bovine cells. First, we demonstrated that the SPRY domain of bovine TRIM5 is the major determinant of anti-viral activity. Second, we found that mutations that allow the capsid to evade rhesus macaque TRIM5α minimally rescued HIV-1 infectivity in bovine-derived MDBK cells. Third, we found that cyclosporine A, which relieves the inhibition of HIV-1 infection in monkey cells, significantly rescued the impaired HIV-1 infectivity in MDBK cells. Lastly, we successfully generated a bovine cell line lacking intact TRIM5 using the CRISPR/Cas9 technique. This TRIM5 knockout cell line displayed significantly higher susceptibility to an HIV-1-based lentiviral vector. In conclusion, our findings provide a promising gene engineering strategy for bovine cells, thereby contributing to innovations in agriculture and improvements in animal health.
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Virucidal activity and mechanism of action of cetylpyridinium chloride against SARS-CoV-2 査読あり
Okamoto N., Saito A., Okabayashi T., Komine A.
Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology 34 ( 6 ) 800 - 804 2022年11月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology
Objective: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the pathogen causing the coronavirus disease 2019 (COVID-19) global pandemic. Recent studies have shown the importance of the throat and salivary glands as sites of virus replication and transmission. The viral host receptor, angiotensin-converting enzyme 2 (ACE2), is broadly enriched in epithelial cells of the salivary glands and oral mucosae. Oral care products containing cetylpyridinium chloride (CPC) as a bactericidal ingredient are known to exhibit antiviral activity against SARS-CoV-2 in vitro. However, the exact mechanism of action remains unknown. Methods: This study examined the antiviral activity of CPC against SARS-CoV-2 and its inhibitory effect on the interaction between the viral spike (S) protein and ACE2 using an enzyme-linked immunosorbent assay. Results: CPC (0.05%, 0.1% and 0.3%) effectively inactivated SARS-CoV-2 within the contact times (20 and 60 s) in directions for use of oral care products in vitro. The binding ability of both the S protein and ACE2 were reduced by CPC. Conclusions: Our results suggest that CPC inhibits the interaction between S protein and ACE2, and thus, reduces infectivity of SARS-CoV-2 and suppresses viral adsorption.
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Survey of hepatitis E virus in pork products and pig stools in Nakhon Pathom Province, Thailand
Thippornchai N., Leaungwutiwong P., Kosoltanapiwat N., Vuong C., Nguyen K., Okabayashi T., Lee A.
Veterinary Medicine and Science 8 ( 5 ) 1975 - 1981 2022年9月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Veterinary Medicine and Science
Background: Hepatitis E virus (HEV) is an important public health threat resulting in more than 3 million symptomatic cases and 70,000 deaths annually. HEV is classified into at least eight genotypes, and five are associated with human infection. Genotypes 1 and 2 primarily affect humans, whereas genotypes 3 and 4 circulate in both humans and swine and are considered zoonotic viruses. Previous studies in Central Thailand have reported human HEV isolates with high similarity to swine strains and high seroprevalence in pigs, suggesting the potential for pig-to-human transmission. Objectives: This study aimed to detect and analyse HEV in pork products and pig stools collected from local markets and pig farms in Nakhon Pathom Province in Central Thailand. Methods: A total of 177 pig stool and 214 pork product samples were detected for HEV by using RT–PCR amplification. Next, nucleotide sequencing and phylogenetic analysis were performed. Results: We found one sample of pork products (1/214, 0.5%), which was a pig liver sample (1/51, 2.0%), and 49 HEV-positive samples in pig stools (49/177, 27.7%). Phylogenetic analysis showed that all these HEV sequences belonged to genotype 3, with a high correlation between our samples and HEV from humans and swine was previously reported in Thailand. Conclusions: This study suggested that the consumption of poorly sanitized or uncooked animal meat or food and frequent exposure to pig stools may be risk factors for HEV infections in humans.
DOI： 10.1002/vms3.854
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Sugamoto K., Tanaka Y.L., Saito A., Goto Y., Nakayama T., Okabayashi T., Kunitake H., Morishita K.
Biochemical and Biophysical Research Communications 615 ( 615 ) 56 - 62 2022年7月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Biochemical and Biophysical Research Communications
With the current worldwide pandemic of COVID-19, there is an urgent need to develop effective treatment and prevention methods against SARS-CoV-2 infection. We have previously reported that the proanthocyanidin (PAC) fraction in blueberry (BB) leaves has strong antiviral activity against hepatitis C virus (HCV) and human T-lymphocytic leukemia virus type 1 (HTLV-1). In this study, we used Kunisato 35 Gou (K35) derived from the rabbit eye blueberry (Vaccinium virgatum Aiton), which has a high PAC content in the leaves and stems. The mean of polymerization (mDP) of PAC in K35 was the highest of 7.88 in Fraction 8 (Fr8) from the stems and 12.28 of Fraction 7 (Fr7) in the leaves. The composition of BB-PAC in K35 is that most are B-type bonds with a small number of A-type bonds and cinchonain I as extension units. A strong antiviral effect was observed in Fr7, with a high polymerized PAC content in both the leaves and stems. Furthermore, when we examined the difference in the action of BB-PAC before and after SARS-CoV-2 infection, we found a stronger inhibitory effect in the pre-infection period. Moreover, BB-PAC Fr7 inhibited the activity of angiotensin II converting enzyme (ACE2), although no effect was observed in a neutralization test of pseudotyped SARS-CoV-2. The viral chymotrypsin-like cysteine protease (3CLpro) of SARS-CoV-2 was also inhibited by BB-PAC Fr7 in leaves and stems. These results indicate that BB-PAC has at least two different inhibitory effects, and that it is effective in suppressing SARS-CoV-2 infection regardless of the time of infection.
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Kaneko C., Saito A., Inagaki H., Sugiyama H., Mazimpaka E., Fujimoto S., Okabayashi T.
Journal of Medical Virology 94 ( 7 ) 3438 - 3441 2022年7月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Journal of Medical Virology
Severe fever with thrombocytopenia syndrome (SFTS) caused by Dabie bandavirus (SFTSV) is a serious public health concern in endemic areas, particularly in Asian and Southeast Asian countries. SFTSV is transmitted by direct contact with body fluids from infected humans and animals. Therefore, environmental hygiene in hospitals and veterinary clinics in SFTSV-endemic areas is highly important. This study assessed the effects of continuous and intermittent irradiation with deep-ultraviolet light-emitting diode (DUV-LED) on SFTSV. Evaluation was performed by conducting plaque assay in which SFTSV irradiated with deep-ultraviolet (DUV; 280 ± 5 nm) was inoculated onto Vero cells. The results showed that continuous and intermittent irradiation for 5 s, resulting in 18.75 mJ/cm2 of cumulative UV exposure, led to a >2.7 and >2.9 log reduction, respectively, corresponding to a >99.8% reduction in infectivity. These results demonstrate that DUV can be utilized for inactivation of SFTSV to maintain environmental hygiene in hospitals and veterinary clinics in endemic countries.
DOI： 10.1002/jmv.27698
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Global Prevalence of Domestic Cat Hepadnavirus: An Emerging Threat to Cats’ Health? 査読あり
Shofa M., Kaneko Y., Takahashi K., Okabayashi T., Saito A.
Frontiers in Microbiology 13 938154 2022年6月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Frontiers in Microbiology
Hepatitis B is an infectious hepatocellular disease of global concern caused by hepatitis B virus (HBV), which belongs to Hepadnaviridae. Recently, a novel HBV-like virus, domestic cat hepadnavirus (DCH), was detected from an immunocompromised cat with a hepatic disease in Australia. Subsequent molecular investigation by independent research groups revealed that its prevalence rates were 6.5% in Australia, 10.8% in Italy, 12.4% in Thailand, 12.3% in Malaysia, 3.08% in the United Kingdom, and 0.78% in Japan. Although the correlation between DCH infection and hepatic diseases remains to be elucidated, understanding the diversity of circulating DCH will contribute to its prevention and control in domestic cats. Herein, we summarize the current epidemiological data of DCH in these countries.
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Kirino Y., Yamanaka A., Ishijima K., Tatemoto K., Maeda K., Okabayashi T.
Journal of Infection and Chemotherapy 28 ( 6 ) 753 - 756 2022年6月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Journal of Infection and Chemotherapy
Introduction: Severe fever with thrombocytopenia syndrome (SFTS) is an emerging tick-born disease and its animal-to-human transmission has come to attention recently. During our sero-survey of SFTS virus (SFTSV) among veterinary professionals in 2018, a veterinarian and his assistant working in an animal hospital were tested positive by enzyme-linked immunosorbent assay (ELISA). An additional survey implied a cluster of SFTS cases in which four more people, a family who brought two sick dogs to the animal hospital in 2003, were involved. This study aimed at assessing the possibility of animal-to-human transmission of SFTSV in this cluster. Methods: Retrospective interviews were performed with the owner family of the dogs and their clinical records were obtained from each hospital. SFTSV-IgG were tested by ELISA and virus neutralization test using the sera collected from them in 2018. Results: The interviews revealed that a total of six people, the two veterinary professionals and the owner family who took care of the sick dogs, suffered from SFTS-like symptoms in the same period of time in 2003. All patients did not have tick bite before the onset and all suspected causative agents were excluded by laboratory tests. The serological tests in this study revealed the four owner family members were all positive for SFTSV antibodies. Conclusions: Considering the extremely low seroprevalence of SFTSV antibodies among inhabitants of the region, the existence of SFTSV antibodies in all these six people presents a possibility that they were involved in an SFTS outbreak originated in the sick dogs in 2003.
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Identification of domestic cat hepadnavirus from a cat blood sample in Japan 査読あり
Takahashi K., Kaneko Y., Shibanai A., Yamamoto S., Katagiri A., Osuga T., Inoue Y., Kuroda K., Tanabe M., Okabayashi T., Naganobu K., Minobe I., Saito A.
Journal of Veterinary Medical Science 84 ( 5 ) 648 - 652 2022年5月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Journal of Veterinary Medical Science
The hepatitis B virus (Hepadnaviridae) induces chronic hepatitis and hepatic cancer in humans. A novel domestic cat hepadnavirus (DCH) was recently identified in several countries, however, the DCH infection status of cats in Japan is unknown. Therefore, we investigated the DCH infection rate of 139 cat samples collected in Japan. We identified one positive blood sample (0.78%) from a 17-year-old female cat with chronically elevated alanine aminotransferase. Phylogenetic analysis demonstrated that the DCH strain identified in this study is genetically different from strains in other countries. Further investigations are required to elucidate the evolution of DCH and the impact of DCH infection on hepatic diseases in domestic cats.
DOI： 10.1292/jvms.22-0010
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Whole-Genome Sequencing of Pasteurella multocida Strain Pm1, Isolated from a Calf
Gondaira S., Fujiki J., Hirano Y., Murata R., Uchida I., Usui M., Iwasaki T., Okabayashi T., Iwano H., Higuchi H.
Microbiology Resource Announcements 11 ( 4 ) e0004222 2022年4月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Microbiology Resource Announcements
Bovine pneumonia is a disease that causes significant economic losses in livestock industries and is vital for animal welfare. The whole-genome sequence of Pasteurella multocida strain Pm1, isolated from a calf suffering from pneumonia in Japan, is reported here.
DOI： 10.1128/mra.00042-22
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Kotaki T., Kurosu T., Grinyo-Escuer A., Davidson E., Churrotin S., Okabayashi T., Puiprom O., Mulyatno K.C., Sucipto T.H., Doranz B.J., Ono K.i., Soegijanto S., Kameoka M.
Scientific Reports 11 ( 1 ) 12987 2021年12月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Scientific Reports
Dengue virus (DENV), from the genus flavivirus of the family flaviviridae, causes serious health problems globally. Human monoclonal antibodies (HuMAb) can be used to elucidate the mechanisms of neutralization and antibody-dependent enhancement (ADE) of DENV infections, leading to the development of a vaccine or therapeutic antibodies. Here, we generated eight HuMAb clones from an Indonesian patient infected with DENV. These HuMAbs exhibited the typical characteristics of weak neutralizing antibodies including high cross-reactivity with other flaviviruses and targeting of the fusion loop epitope (FLE). However, one of the HuMAbs, 3G9, exhibited strong neutralization (NT50 < 0.1 μg/ml) and possessed a high somatic hyper-mutation rate of the variable region, indicating affinity-maturation. Administration of this antibody significantly prolonged the survival of interferon-α/β/γ receptor knockout C57BL/6 mice after a lethal DENV challenge. Additionally, Fc-modified 3G9 that had lost their in vitro ADE activity showed enhanced therapeutic potency in vivo and competed strongly with an ADE-prone antibody in vitro. Taken together, the affinity-matured FLE-targeting antibody 3G9 exhibits promising features for therapeutic application including a low NT50 value, potential for treatment of various kinds of mosquito-borne flavivirus infection, and suppression of ADE. This study demonstrates the therapeutic potency of affinity-matured FLE-targeting antibodies.
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Izzati U.Z., Kaneko Y., Kaneko C., Yoshida A., Suwanruengsri M., Okabayashi T., Hirai T., Yamaguchi R.
Journal of Comparative Pathology 188 44 - 51 2021年10月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Journal of Comparative Pathology
Following isolation of pseudorabies virus (PRV) from two hunting dogs in Oita prefecture, Japan, we investigated the PRV antigen distribution in the tissues of the infected animals. At necropsy, PRV-associated lesions included facial oedema, tonsillar and meningeal congestion, blotchy haemorrhages on the pericardium and mitral valves, and incomplete splenic contraction in one dog, with less prominent findings in the other dog. Multiple pulmonary nodules were seen in both cases, caused by the diploid form of Paragonimus westermani lung flukes, as confirmed by PCR-restriction fragment length polymorphism and gene sequencing analyses. Histological examination revealed that the PRV infection was associated with lesions of non-suppurative encephalitis in the brainstem. PRV antigen was detected in the cerebrum, cerebellum, brainstem, submucosal and myenteric plexuses, and mononuclear cells, mainly in the bone marrow, lymph nodes, tonsils and spleen tissues. There was evidence of PRV dissemination to the brain via the trigeminal or olfactory routes, in addition to possible spread to lymphoid organs via infected mononuclear cells.
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Oba M., Rongduo W., Saito A., Okabayashi T., Yokota T., Yasuoka J., Sato Y., Nishifuji K., Wake H., Nibu Y., Mizutani T.
Biochemical and Biophysical Research Communications 570 21 - 25 2021年9月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Biochemical and Biophysical Research Communications
Natto, a traditional Japanese fermented soybean food, is well known to be nutritious and beneficial for health. In this study, we examined whether natto impairs infection by viruses, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as well as bovine herpesvirus 1 (BHV-1). Interestingly, our results show that both SARS-CoV-2 and BHV-1 treated with a natto extract were fully inhibited infection to the cells. We also found that the glycoprotein D of BHV-1 was shown to be degraded by Western blot analysis and that a recombinant SARS-CoV-2 receptor-binding domain (RBD) was proteolytically degraded when incubated with the natto extract. In addition, RBD protein carrying a point mutation (UK variant N501Y) was also degraded by the natto extract. When the natto extract was heated at 100 °C for 10 min, the ability of both SARS-CoV-2 and BHV-1 to infect to the cells was restored. Consistent with the results of the heat inactivation, a serine protease inhibitor inhibited anti-BHV-1 activity caused by the natto extract. Thus, our findings provide the first evidence that the natto extract contains a protease(s) that inhibits viral infection through the proteolysis of the viral proteins.
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Srijangwad A., Tripipat T., Saeng-chuto K., Jermsujarit P., Tantituvanont A., Okabayashi T., Nilubol D.
Journal of Immunological Methods 494 113045 2021年7月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Journal of Immunological Methods
The objectives of this study are to develop and optimize indirect ELISA based on three coating antigens of porcine epidemic diarrhea virus (PEDV), recombinant spike (S12), nucleocapsid (N), and whole viral (WV) proteins, for the detection of IgG and IgA antibodies in colostrum and milk and to evaluate the diagnostic sensitivity (DSe) and diagnostic specificity (DSp) of the assay as a diagnostic method. Colostrum (n = 347) and milk (n = 272) samples from sows were employed in this assay. Indirect ELISA based on three coating antigens was assessed by receiver operating characteristic (ROC) curve analysis with a virus neutralization (VN) test as a reference method, and the cutoff value for calculating DSe and DSp was determined. S12-ELISA showed higher DSe and DSp of IgG and IgA detection compared to N- and WV-ELISA in both colostrum and milk samples. Moreover, S12-ELISA showed perfect agreement and a high correlation with the VN test, which was better than the N- and WV-ELISA for both IgG and IgA detection in colostrum and milk. In contrast, N-ELISA showed lower DSe and DSp compared to S12- and WV-ELISA, along with a correlation with VN and substantial agreement with the VN test. Nevertheless, our developed ELISAs have accuracy for repeatability in both inter- and intra-assay variation. Overall, this research demonstrates that S12-ELISA is more suitable than WV- and N-ELISA to detect IgG and IgA antibodies against PEDV from both colostrum and milk samples.