Brucella abortus: determination of survival times and evaluation of methods for detection in several matrices
Abstract Background Brucella abortus is a highly pathogenic zoonotic agent, tempting for the development of a rapid diagnostic method to enable adequate treatment and prevent further spread. Enrichment of the bacteria is often used as a first step in diagnostics to increase the bacterial number abov...
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doaj-f8f0f06c501943e2bc3641360c000da62020-11-25T03:52:08ZengBMCBMC Infectious Diseases1471-23342018-06-011811610.1186/s12879-018-3134-5Brucella abortus: determination of survival times and evaluation of methods for detection in several matricesRene Kaden0Sevinc Ferrari1Tomas Jinnerot2Martina Lindberg3Tara Wahab4Moa Lavander5National Veterinary InstituteNational Veterinary InstituteNational Veterinary InstituteNational Veterinary InstituteSwedish Forum for Biopreparedness DiagnosticsSwedish Forum for Biopreparedness DiagnosticsAbstract Background Brucella abortus is a highly pathogenic zoonotic agent, tempting for the development of a rapid diagnostic method to enable adequate treatment and prevent further spread. Enrichment of the bacteria is often used as a first step in diagnostics to increase the bacterial number above the detection limit of the real-time PCR. The enrichment of Brucella spp. takes at least 3 days, which might be avoidable if sensitive PCR methods can be used. Since many matrices contain PCR inhibitors, the limit of detection (LOD) must be determined for each separate matrix. Another aim of this study was the determination of survival of Brucella abortus in the analyzed matrices. Methods The LOD for the detection of B. abortus in 14 matrices, relevant for human medicine, veterinary medicine and food and feed safety, was determined to evaluate the need of a pre-enrichment step prior to real-time PCR. The survival of B. abortus in the spiked matrices was tested by plate count in a 7-day interval for 132 days. Results The limit of detection for B. abortus in most matrices was in the range of 103–104 CFU/g for cultivation and 104–105 CFU/g for direct real-time PCR. The survival time of B. abortus was less than 21 days in apple purée and stomach content and 28 days in water while B. abortus remained viable at day 132 in milk, blood, spinach and minced meat. Conclusions A direct PCR analysis without enrichment of bacteria saves at least 3 days. However, the limit of detection between direct PCR and plate count differs in a 10 fold range. We conclude that this lower sensitivity is acceptable in most cases especially if quick analysis are required.http://link.springer.com/article/10.1186/s12879-018-3134-5Brucella abortusLimit of detectionDiagnosticsSurvival time |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Rene Kaden Sevinc Ferrari Tomas Jinnerot Martina Lindberg Tara Wahab Moa Lavander |
spellingShingle |
Rene Kaden Sevinc Ferrari Tomas Jinnerot Martina Lindberg Tara Wahab Moa Lavander Brucella abortus: determination of survival times and evaluation of methods for detection in several matrices BMC Infectious Diseases Brucella abortus Limit of detection Diagnostics Survival time |
author_facet |
Rene Kaden Sevinc Ferrari Tomas Jinnerot Martina Lindberg Tara Wahab Moa Lavander |
author_sort |
Rene Kaden |
title |
Brucella abortus: determination of survival times and evaluation of methods for detection in several matrices |
title_short |
Brucella abortus: determination of survival times and evaluation of methods for detection in several matrices |
title_full |
Brucella abortus: determination of survival times and evaluation of methods for detection in several matrices |
title_fullStr |
Brucella abortus: determination of survival times and evaluation of methods for detection in several matrices |
title_full_unstemmed |
Brucella abortus: determination of survival times and evaluation of methods for detection in several matrices |
title_sort |
brucella abortus: determination of survival times and evaluation of methods for detection in several matrices |
publisher |
BMC |
series |
BMC Infectious Diseases |
issn |
1471-2334 |
publishDate |
2018-06-01 |
description |
Abstract Background Brucella abortus is a highly pathogenic zoonotic agent, tempting for the development of a rapid diagnostic method to enable adequate treatment and prevent further spread. Enrichment of the bacteria is often used as a first step in diagnostics to increase the bacterial number above the detection limit of the real-time PCR. The enrichment of Brucella spp. takes at least 3 days, which might be avoidable if sensitive PCR methods can be used. Since many matrices contain PCR inhibitors, the limit of detection (LOD) must be determined for each separate matrix. Another aim of this study was the determination of survival of Brucella abortus in the analyzed matrices. Methods The LOD for the detection of B. abortus in 14 matrices, relevant for human medicine, veterinary medicine and food and feed safety, was determined to evaluate the need of a pre-enrichment step prior to real-time PCR. The survival of B. abortus in the spiked matrices was tested by plate count in a 7-day interval for 132 days. Results The limit of detection for B. abortus in most matrices was in the range of 103–104 CFU/g for cultivation and 104–105 CFU/g for direct real-time PCR. The survival time of B. abortus was less than 21 days in apple purée and stomach content and 28 days in water while B. abortus remained viable at day 132 in milk, blood, spinach and minced meat. Conclusions A direct PCR analysis without enrichment of bacteria saves at least 3 days. However, the limit of detection between direct PCR and plate count differs in a 10 fold range. We conclude that this lower sensitivity is acceptable in most cases especially if quick analysis are required. |
topic |
Brucella abortus Limit of detection Diagnostics Survival time |
url |
http://link.springer.com/article/10.1186/s12879-018-3134-5 |
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