Combined Loop-Mediated Isothermal Amplification Assays for Rapid Detection and One-Step Differentiation of Campylobacter jejuni and Campylobacter coli in Meat Products
Copyright © 2021 Kreitlow, Becker, Ahmed, Kittler, Schotte, Plötz and Abdulmawjood..
A loop-mediated isothermal amplification (LAMP) assay system was established, allowing rplD gene-based simultaneous detection of Campylobacter jejuni and Campylobacter coli in enriched meat products. Additionally, one-step differentiation of target species on agar plates was enabled by cdtC gene- and gyrA gene-based duplex LAMP. Both the rplD and cdtC-gyrA LAMP assays amplified the target sequences in all 62 C. jejuni and 27 C. coli strains used for determining inclusivity and revealed 100% exclusivity toward 85 tested non-target species. Throughout the entire experiments, C. jejuni and C. coli strains were 100% distinguishable by melting curves of cdtC and gyrA LAMP products. After 24-h enrichment, the rplD LAMP assay reliably detected initial inoculation levels of 10-100 CFU/g in artificially contaminated minced meat. Investigation of naturally contaminated meat samples revealed a diagnostic accuracy of 95% toward real-time PCR and 94.1% toward the standard culture method applying the 24-h incubation period. Diagnostic sensitivity and specificity, and positive and negative predictive values were 89.8, 100, 100, and 91.2%, respectively, when measured against real-time PCR, and 89.6, 98.1, 97.7, and 91.2%, respectively, when measured against the standard culture method. After 48-h enrichment, the detection limit of the rplD LAMP assay improved to initial inoculation levels of 1-10 CFU/g in artificially contaminated minced meat. Applying the 48-h incubation period on naturally contaminated meat samples resulted in 100% concordant results between rplD LAMP, real-time PCR, and the standard culture method. The established LAMP assay system was proved to be suitable for rapid meat sample screening. Furthermore, it constitutes a promising tool for investigating other Campylobacter sources and could therefore make a valuable contribution to protect consumers from foodborne illness.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2021 |
|---|---|
| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:12 |
|---|---|
| Enthalten in: |
Frontiers in microbiology - 12(2021) vom: 01., Seite 668824 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Kreitlow, Antonia [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
Campylobacter |
|---|
| Anmerkungen: |
Date Revised 29.06.2021 published: Electronic-eCollection Citation Status PubMed-not-MEDLINE |
|---|
| doi: |
10.3389/fmicb.2021.668824 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM32724903X |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM32724903X | ||
| 003 | DE-627 | ||
| 005 | 20231225200348.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231225s2021 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.3389/fmicb.2021.668824 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1090.xml |
| 035 | |a (DE-627)NLM32724903X | ||
| 035 | |a (NLM)34177847 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Kreitlow, Antonia |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Combined Loop-Mediated Isothermal Amplification Assays for Rapid Detection and One-Step Differentiation of Campylobacter jejuni and Campylobacter coli in Meat Products |
| 264 | 1 | |c 2021 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Revised 29.06.2021 | ||
| 500 | |a published: Electronic-eCollection | ||
| 500 | |a Citation Status PubMed-not-MEDLINE | ||
| 520 | |a Copyright © 2021 Kreitlow, Becker, Ahmed, Kittler, Schotte, Plötz and Abdulmawjood. | ||
| 520 | |a A loop-mediated isothermal amplification (LAMP) assay system was established, allowing rplD gene-based simultaneous detection of Campylobacter jejuni and Campylobacter coli in enriched meat products. Additionally, one-step differentiation of target species on agar plates was enabled by cdtC gene- and gyrA gene-based duplex LAMP. Both the rplD and cdtC-gyrA LAMP assays amplified the target sequences in all 62 C. jejuni and 27 C. coli strains used for determining inclusivity and revealed 100% exclusivity toward 85 tested non-target species. Throughout the entire experiments, C. jejuni and C. coli strains were 100% distinguishable by melting curves of cdtC and gyrA LAMP products. After 24-h enrichment, the rplD LAMP assay reliably detected initial inoculation levels of 10-100 CFU/g in artificially contaminated minced meat. Investigation of naturally contaminated meat samples revealed a diagnostic accuracy of 95% toward real-time PCR and 94.1% toward the standard culture method applying the 24-h incubation period. Diagnostic sensitivity and specificity, and positive and negative predictive values were 89.8, 100, 100, and 91.2%, respectively, when measured against real-time PCR, and 89.6, 98.1, 97.7, and 91.2%, respectively, when measured against the standard culture method. After 48-h enrichment, the detection limit of the rplD LAMP assay improved to initial inoculation levels of 1-10 CFU/g in artificially contaminated minced meat. Applying the 48-h incubation period on naturally contaminated meat samples resulted in 100% concordant results between rplD LAMP, real-time PCR, and the standard culture method. The established LAMP assay system was proved to be suitable for rapid meat sample screening. Furthermore, it constitutes a promising tool for investigating other Campylobacter sources and could therefore make a valuable contribution to protect consumers from foodborne illness | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Campylobacter | |
| 650 | 4 | |a LAMP assay | |
| 650 | 4 | |a cdtC gene | |
| 650 | 4 | |a gyrA gene | |
| 650 | 4 | |a meat products | |
| 650 | 4 | |a rplD gene | |
| 700 | 1 | |a Becker, André |e verfasserin |4 aut | |
| 700 | 1 | |a Ahmed, Marwa F E |e verfasserin |4 aut | |
| 700 | 1 | |a Kittler, Sophie |e verfasserin |4 aut | |
| 700 | 1 | |a Schotte, Ulrich |e verfasserin |4 aut | |
| 700 | 1 | |a Plötz, Madeleine |e verfasserin |4 aut | |
| 700 | 1 | |a Abdulmawjood, Amir |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Frontiers in microbiology |d 2010 |g 12(2021) vom: 01., Seite 668824 |w (DE-627)NLM208040617 |x 1664-302X |7 nnns |
| 773 | 1 | 8 | |g volume:12 |g year:2021 |g day:01 |g pages:668824 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.3389/fmicb.2021.668824 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 12 |j 2021 |b 01 |h 668824 | ||