Development of a real-time recombinase-aided amplification assay for rapid and sensitive detection of Edwardsiella piscicida
Edwardsiella piscicida, a significant intracellular pathogen, is widely distributed in aquatic environments and causes systemic infection in various species. Therefore, it’s essential to develop a rapid, uncomplicated and sensitive method for detection of E. piscicida in order to control the transmission of this pathogen effectively. The recombinase-aided amplification (RAA) assay is a newly developed, rapid detection method that has been utilized for various pathogens. In the present study, a real-time RAA (RT-RAA) assay, targeting the conserved positions of the EvpP gene, was successfully established for the detection of E. piscicida. This assay can be performed in a one-step single tube reaction at a temperature of 39°C within 20 min. The RT-RAA assay exhibited a sensitivity of 42 copies per reaction at a 95% probability, which was comparable to the sensitivity of real-time quantitative PCR (qPCR) assay. The specificity assay confirmed that the RT-RAA assay specifically targeted E. piscicida without any cross-reactivity with other important marine bacterial pathogens. Moreover, when clinical specimens were utilized, a perfect agreement of 100% was achieved between the RT-RAA and qPCR assays, resulting a kappa value of 1. These findings indicated that the established RT-RAA assay provided a viable alternative for the rapid, sensitive, and specific detection of E. piscicida..
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2024 |
|---|---|
| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:14 |
|---|---|
| Enthalten in: |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Yuchen Dong [VerfasserIn] |
|---|
| Links: |
doi.org [kostenfrei] |
|---|
| Themen: |
Aquaculture |
|---|
| doi: |
10.3389/fcimb.2024.1355056 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
DOAJ099789736 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | DOAJ099789736 | ||
| 003 | DE-627 | ||
| 005 | 20240414054047.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 240414s2024 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.3389/fcimb.2024.1355056 |2 doi | |
| 035 | |a (DE-627)DOAJ099789736 | ||
| 035 | |a (DE-599)DOAJ0101d7a3ecc848c0ab40fbf457aa6936 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 050 | 0 | |a QR1-502 | |
| 100 | 0 | |a Yuchen Dong |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Development of a real-time recombinase-aided amplification assay for rapid and sensitive detection of Edwardsiella piscicida |
| 264 | 1 | |c 2024 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 520 | |a Edwardsiella piscicida, a significant intracellular pathogen, is widely distributed in aquatic environments and causes systemic infection in various species. Therefore, it’s essential to develop a rapid, uncomplicated and sensitive method for detection of E. piscicida in order to control the transmission of this pathogen effectively. The recombinase-aided amplification (RAA) assay is a newly developed, rapid detection method that has been utilized for various pathogens. In the present study, a real-time RAA (RT-RAA) assay, targeting the conserved positions of the EvpP gene, was successfully established for the detection of E. piscicida. This assay can be performed in a one-step single tube reaction at a temperature of 39°C within 20 min. The RT-RAA assay exhibited a sensitivity of 42 copies per reaction at a 95% probability, which was comparable to the sensitivity of real-time quantitative PCR (qPCR) assay. The specificity assay confirmed that the RT-RAA assay specifically targeted E. piscicida without any cross-reactivity with other important marine bacterial pathogens. Moreover, when clinical specimens were utilized, a perfect agreement of 100% was achieved between the RT-RAA and qPCR assays, resulting a kappa value of 1. These findings indicated that the established RT-RAA assay provided a viable alternative for the rapid, sensitive, and specific detection of E. piscicida. | ||
| 650 | 4 | |a E. piscicida | |
| 650 | 4 | |a aquaculture | |
| 650 | 4 | |a recombinase-aided amplification | |
| 650 | 4 | |a rapid diagnosis | |
| 650 | 4 | |a isothermal amplification | |
| 653 | 0 | |a Microbiology | |
| 700 | 0 | |a Dandan Zhou |e verfasserin |4 aut | |
| 700 | 0 | |a Binzhe Zhang |e verfasserin |4 aut | |
| 700 | 0 | |a Xiaoying Xu |e verfasserin |4 aut | |
| 700 | 0 | |a Jian Zhang |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i In |t Frontiers in Cellular and Infection Microbiology |d Frontiers Media S.A., 2016 |g 14(2024) |w (DE-627)DOAJ000023329 |x 22352988 |7 nnns |
| 773 | 1 | 8 | |g volume:14 |g year:2024 |
| 856 | 4 | 0 | |u https://doi.org/10.3389/fcimb.2024.1355056 |z kostenfrei |
| 856 | 4 | 0 | |u https://doaj.org/article/0101d7a3ecc848c0ab40fbf457aa6936 |z kostenfrei |
| 856 | 4 | 0 | |u https://www.frontiersin.org/articles/10.3389/fcimb.2024.1355056/full |z kostenfrei |
| 856 | 4 | 2 | |u https://doaj.org/toc/2235-2988 |y Journal toc |z kostenfrei |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_DOAJ | ||
| 951 | |a AR | ||
| 952 | |d 14 |j 2024 | ||