%0 Journal Article %K mastitis pathogens %K molecular methods %K conventional bacteriology %K dairy cows %B Berliner und Münchener Tierärztliche Wochenschrift %C Hannover %D 2012 %G English %I Schlütersche Verlagsgesellschaft mbH & Co. KG %P 494-502 %R 10.2376/0005-9366-125-494 %T Mastitis diagnosis in dairy cows using PathoProofTM real-time polymerase chain reaction assay in comparison with conventional bacterial culture in a Northern German field study %V 125 %1 {"oldId":70843,"title":"Mastitis diagnosis in dairy cows using PathoProofTM real-time polymerase chain reaction assay in comparison with conventional bacterial culture in a Northern German field study","teaserText":"In the following field study, the commercial PathoProofTM Mastitis PCR Assay, a real-time PCR for identifying eleven mastitis pathogens and the staphylococcal beta-lactamase gene, was compared with conventional bacterial culture. For this purpose, ...","content":"

Summary<\/span>
In the following field study, the commercial PathoProofTM Mastitis PCR Assay, a real-time PCR for identifying eleven mastitis pathogens and the staphylococcal beta-lactamase gene, was compared with conventional bacterial culture. For this purpose, 681 udder quarter samples from 173 clinically healthy cows with vary- ing somatic cell count from four dairy herds in the region of Osnabr\u00fcck, Lower Saxony, Germany, were collected between July 2010 and February 2011 and sub- jected to PCR and bacterial culture. The frequency of positive pathogen signals was markedly higher with PCR compared with culture (70.6% vs. 32.2%). This was accompanied by a substantial higher percentage of multiple pathogen identi- fications and a lower percentage of single identifications in the PCR compared with bacterial culture. Using bacterial culture as gold standard, moderate to high sensitivities (76.4\u2013100%) and specificities (63.3\u201348.7%) were calculated for six out of seven pathogens with sufficient detection numbers. For Enterococcus spp., the sensitivity was only 4.1%. When the PCR results of pooled udder quarter samples of the 173 cows were compared with the single udder quarter samples, in 72% of the cases, major pathogen DNA was either not found in both types of samples, or in the case of a positive pool sample, the respective pathogens were foundin at least one udder quarter sample. With both methods, the most frequently detected mastitis pathogens were coryneform bacteria (PCR: Corynebacterium bovis), coagulase-negative staphylococci (CNS) and Staphylococcus (S.) aureus, followed by Arcanobacterium pyogenes\/Peptoniphilus indolicus with PCR, and then with both methods, Streptococcus uberis. The staphylococcal beta-lactamase gene was found in 27.7% of the S. aureus and in 37.0% of the CNS identifications.

Keywords:<\/span>
mastitis pathogens, molecular methods, conventional bacteriology, dairy cows


Zusammenfassung<\/span>
In der vorliegenden Feldstudie wurde der kommerzielle PathoProofTM Mastitis PCR Assay, eine real-time PCR zur Identifikation von elf Mastitiserregern und dem Gen f\u00fcr Staphylokokken-Betalaktamase, mit der konventionellen bakteriologischen Untersuchung verglichen. Dazu wurden im Raum Osnabr\u00fcck, Niedersachsen, Deutschland, 681 Viertelgemelksproben von 173 klinisch unauff\u00e4lligen Tierenmit unterschiedlichem somatischem Zellgehalt von vier Milchviehbetrieben im Zeitraum Juli 2010 bis Februar 2011 genommen und kulturell-bakteriologisch sowie mittels PCR untersucht. Die H\u00e4ufigkeit positiver Erregersignale war mitder PCR deutlich hoher als mit der kulturellen Untersuchung (70,6 % vs. 32,2 %). Dies ging einher mit einem wesentlich hoheren Anteil von Mehrfachnachweisen und einem geringeren Anteil von Einzelnachweisen in der PCR im Vergleich zur kulturellen Untersuchung. Unter Ber\u00fccksichtigung der bakteriologischen Untersu- chung als Goldstandard wurden mittlere bis hohe Sensitivit\u00e4ten(76,4\u2013100 %) und Spezifit\u00e4ten (63,3\u201348,7 %) f\u00fcr sechs von sieben Erregern mit ausreichenden Nachweish\u00e4ufigkeiten errechnet. F\u00fcr Enterococcus spp. betrug die Sensitivit\u00e4t nur 4,1 %. Der Vergleich der PCR-Ergebnisse gepoolter Viertelgemelks- proben der 173 K\u00fche mit den einzelnen Viertelergebnissen ergab, dass f\u00fcr \u201emajor pathogens\u201c in 72 % der F\u00e4lle die Erreger-DNS entweder in beiden Probentypen nicht nachweisbar war oder bei einer positiven Poolprobe mindestens eine Viertelgemelksprobe ebenfalls positiv f\u00fcr den jeweiligen \u201emajor pathogen\u201c war. Die am h\u00e4ufigsten nachgewiesenen Mastitiserreger waren f\u00fcr beide Methoden coryneforme Erreger (PCR: Corynebacterium bovis), koagulasenegative Staphy- lokokken (KNS) und Staphylococcus (S.) aureus. An vierter Stelle lag in der PCR Arcanobacterium pyogenes\/Peptoniphilus indolicus. Es folgte wie in der kulturellen Untersuchung Streptococcus uberis. Das Staphylokokken-Betalaktamase-Gen lag bei 27,7 % der S. aureus- und 37.0 % der KNS-Identifikationen vor.

Schl\u00fcsselw\u00f6rter:<\/span>
Mastitiserreger, Molekularbiologie, konventionelle Bakteriologie, Milchk\u00fche<\/p>","categories":["Tier\u00e4rztliche Wochenschrift","Abostufe BMTW","Fachartikel"],"fromDate":"Nov 12, 2012 12:00:00 AM","toDate":"Dec 31, 2050 12:00:00 AM","oldUrls":["http:\/\/vetline.de\/mastitis-pathogens-molecular-methods-conventional-bacteriology-dairy-cows\/150\/3130\/70843"],"doiLanguage":"englisch","doiProductFormat":"Online","doiPublisher":"Schl\u00fctersche Verlagsgesellschaft mbH & Co. KG","doiSerialWorkTitle":"Berl. M\u00fcnch. Tier\u00e4rztl. Wschr.","doiDocumentUri":"http:\/\/www.vetline.de\/mastitis-pathogens-molecular-methods-conventional-bacteriology-dairy-cows\/150\/3130\/70843","doiSource":"Berl. M\u00fcnch. Tier\u00e4rztl. Wschr. 125: 11-12, 494-502 (2012)","doiissn":"0005-9366","doiNr":"10.2376\/0005-9366-125-494","doiFirstPage":"494","doiLastPage":"502","doiTransmitted":true,"doiAuthor":"Spittel S, Hoedemaker M","pdf":{"path":"http:\/\/data\/bmtw_2012_11_0494.pdf","title":"bmtw_2012_11_0494.pdf","description":"Mastitis diagnosis in dairy cows using PathoProofTM real-time polymerase chain reaction assay in comparison with conventional bacterial culture in a Northern German field study

"},"authors":[{"firstName":"S","middleName":"","lastName":"Spittel"},{"firstName":"M","middleName":"","lastName":"Hoedemaker"}],"contentOptimised":"

Summary<\/strong>
In the following field study, the commercial PathoProofTM Mastitis PCR Assay, a real-time PCR for identifying eleven mastitis pathogens and the staphylococcal beta-lactamase gene, was compared with conventional bacterial culture. For this purpose, 681 udder quarter samples from 173 clinically healthy cows with vary- ing somatic cell count from four dairy herds in the region of Osnabr\u00fcck, Lower Saxony, Germany, were collected between July 2010 and February 2011 and sub- jected to PCR and bacterial culture. The frequency of positive pathogen signals was markedly higher with PCR compared with culture (70.6% vs. 32.2%). This was accompanied by a substantial higher percentage of multiple pathogen identi- fications and a lower percentage of single identifications in the PCR compared with bacterial culture. Using bacterial culture as gold standard, moderate to high sensitivities (76.4\u2013100%) and specificities (63.3\u201348.7%) were calculated for six out of seven pathogens with sufficient detection numbers. For Enterococcus spp., the sensitivity was only 4.1%. When the PCR results of pooled udder quarter samples of the 173 cows were compared with the single udder quarter samples, in 72% of the cases, major pathogen DNA was either not found in both types of samples, or in the case of a positive pool sample, the respective pathogens were foundin at least one udder quarter sample. With both methods, the most frequently detected mastitis pathogens were coryneform bacteria (PCR: Corynebacterium bovis), coagulase-negative staphylococci (CNS) and Staphylococcus (S.) aureus, followed by Arcanobacterium pyogenes\/Peptoniphilus indolicus with PCR, and then with both methods, Streptococcus uberis. The staphylococcal beta-lactamase gene was found in 27.7% of the S. aureus and in 37.0% of the CNS identifications.

Keywords:<\/strong>
mastitis pathogens, molecular methods, conventional bacteriology, dairy cows


Zusammenfassung<\/strong>
In der vorliegenden Feldstudie wurde der kommerzielle PathoProofTM Mastitis PCR Assay, eine real-time PCR zur Identifikation von elf Mastitiserregern und dem Gen f\u00fcr Staphylokokken-Betalaktamase, mit der konventionellen bakteriologischen Untersuchung verglichen. Dazu wurden im Raum Osnabr\u00fcck, Niedersachsen, Deutschland, 681 Viertelgemelksproben von 173 klinisch unauff\u00e4lligen Tierenmit unterschiedlichem somatischem Zellgehalt von vier Milchviehbetrieben im Zeitraum Juli 2010 bis Februar 2011 genommen und kulturell-bakteriologisch sowie mittels PCR untersucht. Die H\u00e4ufigkeit positiver Erregersignale war mitder PCR deutlich hoher als mit der kulturellen Untersuchung (70,6 % vs. 32,2 %). Dies ging einher mit einem wesentlich hoheren Anteil von Mehrfachnachweisen und einem geringeren Anteil von Einzelnachweisen in der PCR im Vergleich zur kulturellen Untersuchung. Unter Ber\u00fccksichtigung der bakteriologischen Untersu- chung als Goldstandard wurden mittlere bis hohe Sensitivit\u00e4ten(76,4\u2013100 %) und Spezifit\u00e4ten (63,3\u201348,7 %) f\u00fcr sechs von sieben Erregern mit ausreichenden Nachweish\u00e4ufigkeiten errechnet. F\u00fcr Enterococcus spp. betrug die Sensitivit\u00e4t nur 4,1 %. Der Vergleich der PCR-Ergebnisse gepoolter Viertelgemelks- proben der 173 K\u00fche mit den einzelnen Viertelergebnissen ergab, dass f\u00fcr \u201emajor pathogens\u201c in 72 % der F\u00e4lle die Erreger-DNS entweder in beiden Probentypen nicht nachweisbar war oder bei einer positiven Poolprobe mindestens eine Viertelgemelksprobe ebenfalls positiv f\u00fcr den jeweiligen \u201emajor pathogen\u201c war. Die am h\u00e4ufigsten nachgewiesenen Mastitiserreger waren f\u00fcr beide Methoden coryneforme Erreger (PCR: Corynebacterium bovis), koagulasenegative Staphy- lokokken (KNS) und Staphylococcus (S.) aureus. An vierter Stelle lag in der PCR Arcanobacterium pyogenes\/Peptoniphilus indolicus. Es folgte wie in der kulturellen Untersuchung Streptococcus uberis. Das Staphylokokken-Betalaktamase-Gen lag bei 27,7 % der S. aureus- und 37.0 % der KNS-Identifikationen vor.

Schl\u00fcsselw\u00f6rter:<\/strong>
Mastitiserreger, Molekularbiologie, konventionelle Bakteriologie, Milchk\u00fche<\/p>","primaryLanguage":"englisch","summary":"In the following field study, the commercial PathoProofTM Mastitis PCR Assay, a real-time PCR for identifying eleven mastitis pathogens and the staphylococcal beta-lactamase gene, was compared with conventional bacterial culture. For this purpose, 681 udder quarter samples from 173 clinically healthy cows with vary- ing somatic cell count from four dairy herds in the region of Osnabr\u00fcck, Lower Saxony, Germany, were collected between July 2010 and February 2011 and sub- jected to PCR and bacterial culture. The frequency of positive pathogen signals was markedly higher with PCR compared with culture (70.6% vs. 32.2%). This was accompanied by a substantial higher percentage of multiple pathogen identi- fications and a lower percentage of single identifications in the PCR compared with bacterial culture. Using bacterial culture as gold standard, moderate to high sensitivities (76.4\u2013100%) and specificities (63.3\u201348.7%) were calculated for six out of seven pathogens with sufficient detection numbers. For Enterococcus spp., the sensitivity was only 4.1%. When the PCR results of pooled udder quarter samples of the 173 cows were compared with the single udder quarter samples, in 72% of the cases, major pathogen DNA was either not found in both types of samples, or in the case of a positive pool sample, the respective pathogens were foundin at least one udder quarter sample. With both methods, the most frequently detected mastitis pathogens were coryneform bacteria (PCR: Corynebacterium bovis), coagulase-negative staphylococci (CNS) and Staphylococcus (S.) aureus, followed by Arcanobacterium pyogenes\/Peptoniphilus indolicus with PCR, and then with both methods, Streptococcus uberis. The staphylococcal beta-lactamase gene was found in 27.7% of the S. aureus and in 37.0% of the CNS identifications.","keywords":["mastitis pathogens","molecular methods","conventional bacteriology","dairy cows"],"zusammenfassung":"In der vorliegenden Feldstudie wurde der kommerzielle PathoProofTM Mastitis PCR Assay, eine real-time PCR zur Identifikation von elf Mastitiserregern und dem Gen f\u00fcr Staphylokokken-Betalaktamase, mit der konventionellen bakteriologischen Untersuchung verglichen. Dazu wurden im Raum Osnabr\u00fcck, Niedersachsen, Deutschland, 681 Viertelgemelksproben von 173 klinisch unauff\u00e4lligen Tierenmit unterschiedlichem somatischem Zellgehalt von vier Milchviehbetrieben im Zeitraum Juli 2010 bis Februar 2011 genommen und kulturell-bakteriologisch sowie mittels PCR untersucht. Die H\u00e4ufigkeit positiver Erregersignale war mitder PCR deutlich hoher als mit der kulturellen Untersuchung (70,6 % vs. 32,2 %). Dies ging einher mit einem wesentlich hoheren Anteil von Mehrfachnachweisen und einem geringeren Anteil von Einzelnachweisen in der PCR im Vergleich zur kulturellen Untersuchung. Unter Ber\u00fccksichtigung der bakteriologischen Untersu- chung als Goldstandard wurden mittlere bis hohe Sensitivit\u00e4ten(76,4\u2013100 %) und Spezifit\u00e4ten (63,3\u201348,7 %) f\u00fcr sechs von sieben Erregern mit ausreichenden Nachweish\u00e4ufigkeiten errechnet. F\u00fcr Enterococcus spp. betrug die Sensitivit\u00e4t nur 4,1 %. Der Vergleich der PCR-Ergebnisse gepoolter Viertelgemelks- proben der 173 K\u00fche mit den einzelnen Viertelergebnissen ergab, dass f\u00fcr \u201emajor pathogens\u201c in 72 % der F\u00e4lle die Erreger-DNS entweder in beiden Probentypen nicht nachweisbar war oder bei einer positiven Poolprobe mindestens eine Viertelgemelksprobe ebenfalls positiv f\u00fcr den jeweiligen \u201emajor pathogen\u201c war. Die am h\u00e4ufigsten nachgewiesenen Mastitiserreger waren f\u00fcr beide Methoden coryneforme Erreger (PCR: Corynebacterium bovis), koagulasenegative Staphy- lokokken (KNS) und Staphylococcus (S.) aureus. An vierter Stelle lag in der PCR Arcanobacterium pyogenes\/Peptoniphilus indolicus. Es folgte wie in der kulturellen Untersuchung Streptococcus uberis. Das Staphylokokken-Betalaktamase-Gen lag bei 27,7 % der S. aureus- und 37.0 % der KNS-Identifikationen vor.","schluesselwoerter":["Mastitiserreger","Molekularbiologie","konventionelle Bakteriologie","Milchk\u00fche"],"translatedTitle":"In the following field study, the commercial PathoProofTM Mastitis PCR Assay, a real-time PCR for identifying eleven mastitis pathogens and the staphylococcal beta-lactamase gene, was compared with conventional bacterial culture. For this purpose, ...","abstractE":"In the following field study, the commercial PathoProofTM Mastitis PCR Assay, a real-time PCR for identifying eleven mastitis pathogens and the staphylococcal beta-lactamase gene, was compared with conventional bacterial culture. For this purpose, 681 udder quarter samples from 173 clinically healthy cows with vary- ing somatic cell count from four dairy herds in the region of Osnabr\u00fcck, Lower Saxony, Germany, were collected between July 2010 and February 2011 and sub- jected to PCR and bacterial culture. The frequency of positive pathogen signals was markedly higher with PCR compared with culture (70.6% vs. 32.2%). This was accompanied by a substantial higher percentage of multiple pathogen identi- fications and a lower percentage of single identifications in the PCR compared with bacterial culture. Using bacterial culture as gold standard, moderate to high sensitivities (76.4\u2013100%) and specificities (63.3\u201348.7%) were calculated for six out of seven pathogens with sufficient detection numbers. For Enterococcus spp., the sensitivity was only 4.1%. When the PCR results of pooled udder quarter samples of the 173 cows were compared with the single udder quarter samples, in 72% of the cases, major pathogen DNA was either not found in both types of samples, or in the case of a positive pool sample, the respective pathogens were foundin at least one udder quarter sample. With both methods, the most frequently detected mastitis pathogens were coryneform bacteria (PCR: Corynebacterium bovis), coagulase-negative staphylococci (CNS) and Staphylococcus (S.) aureus, followed by Arcanobacterium pyogenes\/Peptoniphilus indolicus with PCR, and then with both methods, Streptococcus uberis. The staphylococcal beta-lactamase gene was found in 27.7% of the S. aureus and in 37.0% of the CNS identifications.","date":{"year":2012,"date":"11\/2012","accepted":"2012-11-12"},"volume":"125","openAccess":false,"journal":"Berliner und M\u00fcnchener Tier\u00e4rztliche Wochenschrift","titleImageId":944,"pages":"494-502","redirects":["mastitis-pathogens-molecular-methods-conventional-bacteriology-dairy-cows\/150\/3130\/70843"],"tierartCategories":[],"artikelartCategories":["Tier\u00e4rztliche Wochenschrift","Abostufe BMTW","Fachartikel"]} %8 11/2012