The Development of a One-Step PCR Assay for Rapid Detection of an Attenuated Vaccine Strain of Duck Hepatitis Virus Type 3 in Korea
Duck hepatitis A virus type 3 (DHAV-3) is a viral pathogen that causes acute, high-mortality hepatitis in ducklings, and vaccination with attenuated live vaccines is currently the main preventive measure against it. However, differentiating infected from vaccinated animals (DIVA) is crucial for clin...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-12-01
|
| Series: | Veterinary Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2306-7381/12/1/8 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832587338048339968 |
|---|---|
| author | Cheng-Dong Yu Jong-Yeol Park Sang-Won Kim Yu-Ri Choi Se-Yeoun Cha Hyung-Kwan Jang Min Kang Bai Wei |
| author_facet | Cheng-Dong Yu Jong-Yeol Park Sang-Won Kim Yu-Ri Choi Se-Yeoun Cha Hyung-Kwan Jang Min Kang Bai Wei |
| author_sort | Cheng-Dong Yu |
| collection | DOAJ |
| description | Duck hepatitis A virus type 3 (DHAV-3) is a viral pathogen that causes acute, high-mortality hepatitis in ducklings, and vaccination with attenuated live vaccines is currently the main preventive measure against it. However, differentiating infected from vaccinated animals (DIVA) is crucial for clinical diagnosis and effective disease control. This study aimed to develop a rapid mismatch amplification mutation assay PCR (MAMA-PCR) diagnostic method to simultaneously detect and differentiate between wild-type and vaccine strains. The method was specifically designed to target the critical single-nucleotide polymorphism (SNP) site (T→C at position 1143 in the VP0 gene) unique to the Korean vaccine strain AP04203-P100. MAMA-PCR demonstrated high sensitivity and specificity, with detection limits as low as 10<sup>2.4</sup> ELD<sub>50</sub>/mL for wild strains and 10<sup>0.5</sup> ELD<sub>50</sub>/mL for vaccine strains, and showed no cross-reactivity with 11 other common duck pathogens. The clinical sample results were completely consistent with those obtained using nested PCR detection and gold-standard sequencing. In summary, we successfully developed a rapid, one-step MAMA-PCR method that is more suitable for clinical diagnosis than traditional sequencing methods. |
| format | Article |
| id | doaj-art-16021913cb08480bbe5e055fcf369af1 |
| institution | Kabale University |
| issn | 2306-7381 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj-art-16021913cb08480bbe5e055fcf369af12025-01-24T13:51:58ZengMDPI AGVeterinary Sciences2306-73812024-12-01121810.3390/vetsci12010008The Development of a One-Step PCR Assay for Rapid Detection of an Attenuated Vaccine Strain of Duck Hepatitis Virus Type 3 in KoreaCheng-Dong Yu0Jong-Yeol Park1Sang-Won Kim2Yu-Ri Choi3Se-Yeoun Cha4Hyung-Kwan Jang5Min Kang6Bai Wei7Department of Avian Diseases, College of Veterinary Medicine and Center for Avian Disease, Jeonbuk National University, Iksan 54596, Republic of KoreaDepartment of Avian Diseases, College of Veterinary Medicine and Center for Avian Disease, Jeonbuk National University, Iksan 54596, Republic of KoreaDepartment of Avian Diseases, College of Veterinary Medicine and Center for Avian Disease, Jeonbuk National University, Iksan 54596, Republic of KoreaDepartment of Avian Diseases, College of Veterinary Medicine and Center for Avian Disease, Jeonbuk National University, Iksan 54596, Republic of KoreaDepartment of Avian Diseases, College of Veterinary Medicine and Center for Avian Disease, Jeonbuk National University, Iksan 54596, Republic of KoreaDepartment of Avian Diseases, College of Veterinary Medicine and Center for Avian Disease, Jeonbuk National University, Iksan 54596, Republic of KoreaDepartment of Avian Diseases, College of Veterinary Medicine and Center for Avian Disease, Jeonbuk National University, Iksan 54596, Republic of KoreaDepartment of Avian Diseases, College of Veterinary Medicine and Center for Avian Disease, Jeonbuk National University, Iksan 54596, Republic of KoreaDuck hepatitis A virus type 3 (DHAV-3) is a viral pathogen that causes acute, high-mortality hepatitis in ducklings, and vaccination with attenuated live vaccines is currently the main preventive measure against it. However, differentiating infected from vaccinated animals (DIVA) is crucial for clinical diagnosis and effective disease control. This study aimed to develop a rapid mismatch amplification mutation assay PCR (MAMA-PCR) diagnostic method to simultaneously detect and differentiate between wild-type and vaccine strains. The method was specifically designed to target the critical single-nucleotide polymorphism (SNP) site (T→C at position 1143 in the VP0 gene) unique to the Korean vaccine strain AP04203-P100. MAMA-PCR demonstrated high sensitivity and specificity, with detection limits as low as 10<sup>2.4</sup> ELD<sub>50</sub>/mL for wild strains and 10<sup>0.5</sup> ELD<sub>50</sub>/mL for vaccine strains, and showed no cross-reactivity with 11 other common duck pathogens. The clinical sample results were completely consistent with those obtained using nested PCR detection and gold-standard sequencing. In summary, we successfully developed a rapid, one-step MAMA-PCR method that is more suitable for clinical diagnosis than traditional sequencing methods.https://www.mdpi.com/2306-7381/12/1/8duck hepatitis A virus type 3differentiating infected from vaccinated animalsmismatch amplification mutation assayrapid diagnostic |
| spellingShingle | Cheng-Dong Yu Jong-Yeol Park Sang-Won Kim Yu-Ri Choi Se-Yeoun Cha Hyung-Kwan Jang Min Kang Bai Wei The Development of a One-Step PCR Assay for Rapid Detection of an Attenuated Vaccine Strain of Duck Hepatitis Virus Type 3 in Korea Veterinary Sciences duck hepatitis A virus type 3 differentiating infected from vaccinated animals mismatch amplification mutation assay rapid diagnostic |
| title | The Development of a One-Step PCR Assay for Rapid Detection of an Attenuated Vaccine Strain of Duck Hepatitis Virus Type 3 in Korea |
| title_full | The Development of a One-Step PCR Assay for Rapid Detection of an Attenuated Vaccine Strain of Duck Hepatitis Virus Type 3 in Korea |
| title_fullStr | The Development of a One-Step PCR Assay for Rapid Detection of an Attenuated Vaccine Strain of Duck Hepatitis Virus Type 3 in Korea |
| title_full_unstemmed | The Development of a One-Step PCR Assay for Rapid Detection of an Attenuated Vaccine Strain of Duck Hepatitis Virus Type 3 in Korea |
| title_short | The Development of a One-Step PCR Assay for Rapid Detection of an Attenuated Vaccine Strain of Duck Hepatitis Virus Type 3 in Korea |
| title_sort | development of a one step pcr assay for rapid detection of an attenuated vaccine strain of duck hepatitis virus type 3 in korea |
| topic | duck hepatitis A virus type 3 differentiating infected from vaccinated animals mismatch amplification mutation assay rapid diagnostic |
| url | https://www.mdpi.com/2306-7381/12/1/8 |
| work_keys_str_mv | AT chengdongyu thedevelopmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT jongyeolpark thedevelopmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT sangwonkim thedevelopmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT yurichoi thedevelopmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT seyeouncha thedevelopmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT hyungkwanjang thedevelopmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT minkang thedevelopmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT baiwei thedevelopmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT chengdongyu developmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT jongyeolpark developmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT sangwonkim developmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT yurichoi developmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT seyeouncha developmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT hyungkwanjang developmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT minkang developmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea AT baiwei developmentofaonesteppcrassayforrapiddetectionofanattenuatedvaccinestrainofduckhepatitisvirustype3inkorea |