A sensitive, specific and simple loop mediated isothermal amplification method for rapid detection of campylobacter spp. In broiler production

Quyen Than Linh, Steen Nordentoft, Vinayaka Aaydha Chidambara, Tien Ngo Anh, Pia Engelsmann, Yi Sun, Mogens Madsen, Dang Duong Bang*, Anders Wolff

*Corresponding author for this work

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Abstract

Campylobacteriosis is one of the most common foodborne diseases worldwide. Two Campylobacter species – C. jejuni and C. coli in poultry and poultry products are considered to be the main source of human campylobacteriosis. Therefore, studying Campylobacter status in poultry flocks is needed to prevent transmission of disease and reduce human risk, health cost, and economic losses. In this study, we adapted and used a Loop-Mediated Isothermal Amplification (LAMP) assay for specific, sensitive, simple and cost-effective rapid detection of C. jejuni and C. coli in the poultry production chain. Amplified LAMP products were detected using a small, low-cost portable commercial blue LED transilluminator and a direct visual detection strategy was demonstrated. By using optimized conditions for amplification a limit of detection (LOD) of 50 CFU/ml was achieved for testing of C. jejuni and C. coli in spiked chicken feces without enrichment. The method took 60–70 min from receiving the samples to the final results (including 30 min for amplification). The optimized LAMP showed a relative accuracy of 98.4%, a specificity of 97.9%, and a sensitivity of 100% in comparison to real-time PCR method. Cohen’s kappa index also showed an excellent agreement (0.94) between the two methods. The results showed that the method is specific, sensitive and is suitable to develop for rapid detection of Campylobacter spp. at poultry production.

Original languageEnglish
Article number2443
JournalFrontiers in Microbiology
Volume10
Number of pages10
ISSN1664-302X
DOIs
Publication statusPublished - 2019

Keywords

  • Broiler chicken production
  • Broiler fecal sample
  • Campylobacter spp
  • Campylobacteriosis
  • Loop mediated isothermal amplification
  • Rapid detection

Cite this

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title = "A sensitive, specific and simple loop mediated isothermal amplification method for rapid detection of campylobacter spp. In broiler production",
abstract = "Campylobacteriosis is one of the most common foodborne diseases worldwide. Two Campylobacter species – C. jejuni and C. coli in poultry and poultry products are considered to be the main source of human campylobacteriosis. Therefore, studying Campylobacter status in poultry flocks is needed to prevent transmission of disease and reduce human risk, health cost, and economic losses. In this study, we adapted and used a Loop-Mediated Isothermal Amplification (LAMP) assay for specific, sensitive, simple and cost-effective rapid detection of C. jejuni and C. coli in the poultry production chain. Amplified LAMP products were detected using a small, low-cost portable commercial blue LED transilluminator and a direct visual detection strategy was demonstrated. By using optimized conditions for amplification a limit of detection (LOD) of 50 CFU/ml was achieved for testing of C. jejuni and C. coli in spiked chicken feces without enrichment. The method took 60–70 min from receiving the samples to the final results (including 30 min for amplification). The optimized LAMP showed a relative accuracy of 98.4{\%}, a specificity of 97.9{\%}, and a sensitivity of 100{\%} in comparison to real-time PCR method. Cohen’s kappa index also showed an excellent agreement (0.94) between the two methods. The results showed that the method is specific, sensitive and is suitable to develop for rapid detection of Campylobacter spp. at poultry production.",
keywords = "Broiler chicken production, Broiler fecal sample, Campylobacter spp, Campylobacteriosis, Loop mediated isothermal amplification, Rapid detection",
author = "{Than Linh}, Quyen and Steen Nordentoft and {Aaydha Chidambara}, Vinayaka and Anh, {Tien Ngo} and Pia Engelsmann and Yi Sun and Mogens Madsen and Bang, {Dang Duong} and Anders Wolff",
year = "2019",
doi = "10.3389/fmicb.2019.02443",
language = "English",
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journal = "Frontiers in Microbiology",
issn = "1664-302X",
publisher = "Frontiers Media S.A.",

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TY - JOUR

T1 - A sensitive, specific and simple loop mediated isothermal amplification method for rapid detection of campylobacter spp. In broiler production

AU - Than Linh, Quyen

AU - Nordentoft, Steen

AU - Aaydha Chidambara, Vinayaka

AU - Anh, Tien Ngo

AU - Engelsmann, Pia

AU - Sun, Yi

AU - Madsen, Mogens

AU - Bang, Dang Duong

AU - Wolff, Anders

PY - 2019

Y1 - 2019

N2 - Campylobacteriosis is one of the most common foodborne diseases worldwide. Two Campylobacter species – C. jejuni and C. coli in poultry and poultry products are considered to be the main source of human campylobacteriosis. Therefore, studying Campylobacter status in poultry flocks is needed to prevent transmission of disease and reduce human risk, health cost, and economic losses. In this study, we adapted and used a Loop-Mediated Isothermal Amplification (LAMP) assay for specific, sensitive, simple and cost-effective rapid detection of C. jejuni and C. coli in the poultry production chain. Amplified LAMP products were detected using a small, low-cost portable commercial blue LED transilluminator and a direct visual detection strategy was demonstrated. By using optimized conditions for amplification a limit of detection (LOD) of 50 CFU/ml was achieved for testing of C. jejuni and C. coli in spiked chicken feces without enrichment. The method took 60–70 min from receiving the samples to the final results (including 30 min for amplification). The optimized LAMP showed a relative accuracy of 98.4%, a specificity of 97.9%, and a sensitivity of 100% in comparison to real-time PCR method. Cohen’s kappa index also showed an excellent agreement (0.94) between the two methods. The results showed that the method is specific, sensitive and is suitable to develop for rapid detection of Campylobacter spp. at poultry production.

AB - Campylobacteriosis is one of the most common foodborne diseases worldwide. Two Campylobacter species – C. jejuni and C. coli in poultry and poultry products are considered to be the main source of human campylobacteriosis. Therefore, studying Campylobacter status in poultry flocks is needed to prevent transmission of disease and reduce human risk, health cost, and economic losses. In this study, we adapted and used a Loop-Mediated Isothermal Amplification (LAMP) assay for specific, sensitive, simple and cost-effective rapid detection of C. jejuni and C. coli in the poultry production chain. Amplified LAMP products were detected using a small, low-cost portable commercial blue LED transilluminator and a direct visual detection strategy was demonstrated. By using optimized conditions for amplification a limit of detection (LOD) of 50 CFU/ml was achieved for testing of C. jejuni and C. coli in spiked chicken feces without enrichment. The method took 60–70 min from receiving the samples to the final results (including 30 min for amplification). The optimized LAMP showed a relative accuracy of 98.4%, a specificity of 97.9%, and a sensitivity of 100% in comparison to real-time PCR method. Cohen’s kappa index also showed an excellent agreement (0.94) between the two methods. The results showed that the method is specific, sensitive and is suitable to develop for rapid detection of Campylobacter spp. at poultry production.

KW - Broiler chicken production

KW - Broiler fecal sample

KW - Campylobacter spp

KW - Campylobacteriosis

KW - Loop mediated isothermal amplification

KW - Rapid detection

U2 - 10.3389/fmicb.2019.02443

DO - 10.3389/fmicb.2019.02443

M3 - Journal article

C2 - 31708907

AN - SCOPUS:85074583754

VL - 10

JO - Frontiers in Microbiology

JF - Frontiers in Microbiology

SN - 1664-302X

M1 - 2443

ER -