TY - JOUR KW - Salmonella AU - S Braun AU - U Methner AB - There is a strong interest to reduce the expenditure for the detection of Salmonella spp. from animal faeces and environmental samples from primary production according to ISO 6519:2002 Annex D by including a rapid and effective method to detect Salmonella spp. already after pre-enrichment in BPW. It has been shown that real-time PCR methods are very effective to detect Salmonella organisms after pre-enrichment of foods. However, materials from primary animal production compose of much higher amounts of substances which might inhibit the sensitivity of real-time PCR. Different techniques of DNA isolation after preenrichment of artificially inoculated bovine faecal material were used to compare their detection limit and detection probability using an invA 5’ nuclease real-time PCR approach. A detection probability of 100% was shown at 105 cfu/ml using the QIAamp® DNA Stool Mini Kit (Qiagen, Germany), at 104 cfu/ml using the High Pure PCR Template Preparation Kit® (Roche, Germany) and at 103 cfu/ml using thermal cell lysis or an in-house lab protocol, respectively. In comparison DNA isolation by thermal cell lysis revealed a very good detection limit, low costs and almost no risks of contamination. Furthermore, caecal contents from pigs were analysed by ISO 6519:2002 Annex D and the invA real-time PCR using thermal cell lysis for DNA extraction. As a result neither false positive nor false negative findings were obtained. Inclusion of the real-time PCR after pre-enrichment of samples in BPW followed by bacterial detection of Salmonella only with samples positive with real-time PCR might be a valuable tool to fulfil the international standard of ISO 6519:2002 Annex D but also to diminish the expenditures. However, it must be stated that the modification of an international standard method and its use in routine diagnostic requires the validation and registration of national and/ or international competent authorities. BT - Berliner und Münchener Tierärztliche Wochenschrift C1 - {"oldId":69746,"title":"Comparison of DNA isolation methods and detection of Salmonella spp. from animal faeces and dust using invA realtime PCR","topline":"","teaserText":"","content":"

Summary<\/span>
\r\n\r\nThere is a strong interest to reduce the expenditure for the detection of Salmonella<\/span>\r\nspp. from animal faeces and environmental samples from primary production\r\naccording to ISO 6519:2002 Annex D by including a rapid and effective\r\nmethod to detect Salmonella<\/span> spp. already after pre-enrichment in BPW. It has\r\nbeen shown that real-time PCR methods are very effective to detect Salmonella<\/span>\r\norganisms after pre-enrichment of foods. However, materials from primary animal\r\nproduction compose of much higher amounts of substances which might inhibit\r\nthe sensitivity of real-time PCR. Different techniques of DNA isolation after preenrichment\r\nof artificially inoculated bovine faecal material were used to compare\r\ntheir detection limit and detection probability using an invA<\/span> 5\u0092 nuclease real-time\r\nPCR approach. A detection probability of 100% was shown at 105<\/span> cfu\/ml using\r\nthe QIAamp\u00ae DNA Stool Mini Kit (Qiagen, Germany), at 104<\/span> cfu\/ml using the High\r\nPure PCR Template Preparation Kit\u00ae (Roche, Germany) and at 103<\/span> cfu\/ml using\r\nthermal cell lysis or an in-house lab protocol, respectively. In comparison DNA\r\nisolation by thermal cell lysis revealed a very good detection limit, low costs and\r\nalmost no risks of contamination. Furthermore, caecal contents from pigs were\r\nanalysed by ISO 6519:2002 Annex D and the invA<\/span> real-time PCR using thermal\r\ncell lysis for DNA extraction. As a result neither false positive nor false negative\r\nfindings were obtained. Inclusion of the real-time PCR after pre-enrichment of\r\nsamples in BPW followed by bacterial detection of Salmonella<\/span> only with samples\r\npositive with real-time PCR might be a valuable tool to fulfil the international\r\nstandard of ISO 6519:2002 Annex D but also to diminish the expenditures.\r\nHowever, it must be stated that the modification of an international standard\r\nmethod and its use in routine diagnostic requires the validation and registration\r\nof national and\/ or international competent authorities.\r\n\r\n

\r\nKeywords:<\/span>
\r\n\r\n\r\nSalmonella<\/span>, polymerase-chain-reaction, DNA isolation, faecal samples,\r\nISO 6519 Annex D\r\n\r\n\r\n\r\n


\r\nZusammenfassung<\/span>
\r\n\r\n\r\nEs gibt ein starkes Interesse, den Material- und Zeitaufwand f\u00fcr den Nachweis von\r\nSalmonella<\/span> spp. aus Tierkot und Umgebungsproben entsprechend der ISO Norm\r\n6519:2002 Anhang D durch die Einbeziehung einer schnellen und effektiven\r\nMethode zum Nachweis von Salmonellen direkt nach der Voranreicherung zu verringern.\r\nEs wurde gezeigt, dass die real-time-PCR eine sehr effektive Methode f\u00fcr\r\nden Nachweis von Salmonellen aus Lebensmitteln nach einer Voranreicherung\r\nist. Im Gegensatz dazu sind in Proben aus dem Tierbereich jedoch viel gr\u00f6\u00dfere\r\nMengen an bakterieller Sekund\u00e4rflora und an hemmenden Substanzen vorhanden,\r\ndie die Sensitivit\u00e4t der real-time-PCR beeintr\u00e4chtigen k\u00f6nnen.\r\nDaher wurden verschiedene Methoden zur Isolation von DNA nach einer Voranreicherung\r\nvon Salmonellen mit Tierkot verglichen, um das Detektionslimit und\r\ndie Detektionswahrscheinlichkeit mithilfe einer invA<\/span>-basierten real-time-PCR\r\nzu bestimmen. Eine Detektionswahrscheinlichkeit von 100 % konnte mit dem\r\nQIAamp\u00ae DNA Stool Mini Kit (Qiagen, Deutschland) erst ab 105<\/span> kbE\/ml, mit dem\r\nHigh Pure PCR Template Preparation Kit\u00ae (Roche, Deutschland) ab 104<\/span> kbE\/ml und\r\nsowohl bei der Kochlyse als auch bei einem neu entwickelten Hausprotokoll ab\r\n103<\/span> kbE\/ml erreicht werden. Im Vergleich aller verwendeten DNA-Extraktionsmethoden\r\nbesitzt die Kochlyse eine sehr gute Nachweisgrenze, ein geringes Kontaminationsrisiko\r\nund verursacht sehr geringe Kosten. Dar\u00fcber hinaus wurden\r\nProben aus dem Z\u00e4kum-Inhalt von Mastschweinen nach ISO 6519:2002 Annex D\r\nund der invA<\/span>-basierten real-time-PCR mit Kochlyse zur DNA-Isolation untersucht.\r\nDer Vergleich zeigte, dass weder falsch positive noch falsch negative Ergebnisse\r\nauftraten. Die Einbeziehung der invA<\/span>-real-time-PCR nach einer Voranreicherung\r\nder Proben und die nachfolgende Fortsetzung des Nachweises von Salmonellen\r\nnur mit den Proben, die mittels real-time-PCR positiv waren, k\u00f6nnte ein Verfahren\r\ndarstellen, um sowohl die Anforderungen der ISO 6519:2002 Anhang D zu erf\u00fcllen\r\naber gleicherma\u00dfen den Aufwand f\u00fcr diese Methode zu verringen. Es muss\r\njedoch darauf hingewiesen werden, dass die Modifikation einer international\r\ng\u00fcltigen Standardmethode und deren Anwendung in der Routinediagnostik eine\r\nentsprechende Validierung und Zulassung durch die zust\u00e4ndigen nationalen und\r\ninternationalen Beh\u00f6rden erfordert.\r\n\r\n\r\n\r\n

\r\nSchl\u00fcsselw\u00f6rter:<\/span>
\r\n\r\nSalmonella<\/span>, Polymerasekettenreaktion, DNS-Isolierung, Tierkot,\r\nISO 6519 Anhang D\r\n\r\n\r\n\r\n\r\n\r\n


\r\n\r\n\r\n\r\n\r\n\r\n\r\n\r\n\r\nQuelle:
Berl. M\u00fcnch. Tier\u00e4rztl. Wschr. 124: 5-6, 177-185 (2011)
\r\nAutor: Braun SD, Methner U
\r\nDOI-Nummer: 10.2376\/0005-9366-124-177

\r\n\u00a9 Schl\u00fctersche Verlagsgesellschaft mbH amp; Co. KG \r\n\r\n\r\n\r\n\r\n\r\n <\/a><\/p>

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<\/p>","categories":["Open Access","Tier\u00e4rztliche Wochenschrift","Abostufe BMTW","Fachartikel","Abostufe frei"],"fromDate":"Apr 19, 2011 12:00:00 AM","toDate":"Dec 31, 2050 12:00:00 AM","oldUrls":["http:\/\/vetline.de\/open-access-salmonellai-polymerase-chain-reaction-dna-isolation-faecal-samples\/150\/3216\/69746","http:\/\/vetline.de\/open-access-salmonellai-polymerase-chain-reaction-dna-isolation-faecal-samples\/150\/3130\/69746"],"doiLanguage":"englisch","doiProductFormat":"Online","doiPublisher":"Schl\u00fctersche Verlagsgesellschaft mbH & Co. KG","doiSerialWorkTitle":"Berl. M\u00fcnch. Tier\u00e4rztl. Wschr.","doiDocumentUri":"http:\/\/www.vetline.de\/open-access-salmonellai-polymerase-chain-reaction-dna-isolation-faecal-samples\/150\/3130\/69746","doiSource":"Berl. M\u00fcnch. Tier\u00e4rztl. Wschr. 124: 5-6, 177-185 (2011)","doiissn":"0005-9366","doiNr":"10.2376\/0005-9366-124-177","doiFirstPage":"177","doiLastPage":"185","doiTransmitted":true,"doiAuthor":"Braun SD, Methner U","pdf":{"path":"http:\/\/data\/bmtw_2011_05_0177.pdf","title":"bmtw - Comparison of DNA isolation methods and detection of Salmonella spp. from animal faeces and dust using invA realtime PCR ","description":""},"authors":[{"firstName":"S","middleName":"D","lastName":"Braun"},{"firstName":"U","middleName":"","lastName":"Methner"}],"contentOptimised":"

Summary<\/strong>
\n\nThere is a strong interest to reduce the expenditure for the detection of Salmonella<\/strong>\nspp. from animal faeces and environmental samples from primary production\naccording to ISO 6519:2002 Annex D by including a rapid and effective\nmethod to detect Salmonella<\/strong> spp. already after pre-enrichment in BPW. It has\nbeen shown that real-time PCR methods are very effective to detect Salmonella<\/strong>\norganisms after pre-enrichment of foods. However, materials from primary animal\nproduction compose of much higher amounts of substances which might inhibit\nthe sensitivity of real-time PCR. Different techniques of DNA isolation after preenrichment\nof artificially inoculated bovine faecal material were used to compare\ntheir detection limit and detection probability using an invA<\/strong> 5\u2019 nuclease real-time\nPCR approach. A detection probability of 100% was shown at 105 cfu\/ml using\nthe QIAamp\u00ae DNA Stool Mini Kit (Qiagen, Germany), at 104 cfu\/ml using the High\nPure PCR Template Preparation Kit\u00ae (Roche, Germany) and at 103 cfu\/ml using\nthermal cell lysis or an in-house lab protocol, respectively. In comparison DNA\nisolation by thermal cell lysis revealed a very good detection limit, low costs and\nalmost no risks of contamination. Furthermore, caecal contents from pigs were\nanalysed by ISO 6519:2002 Annex D and the invA<\/strong> real-time PCR using thermal\ncell lysis for DNA extraction. As a result neither false positive nor false negative\nfindings were obtained. Inclusion of the real-time PCR after pre-enrichment of\nsamples in BPW followed by bacterial detection of Salmonella<\/strong> only with samples\npositive with real-time PCR might be a valuable tool to fulfil the international\nstandard of ISO 6519:2002 Annex D but also to diminish the expenditures.\nHowever, it must be stated that the modification of an international standard\nmethod and its use in routine diagnostic requires the validation and registration\nof national and\/ or international competent authorities.\n\n

\nKeywords:<\/strong>
\n\n\nSalmonella<\/strong>, polymerase-chain-reaction, DNA isolation, faecal samples,\nISO 6519 Annex D\n\n\n\n


\nZusammenfassung<\/strong>
\n\n\nEs gibt ein starkes Interesse, den Material- und Zeitaufwand f\u00fcr den Nachweis von\nSalmonella<\/strong> spp. aus Tierkot und Umgebungsproben entsprechend der ISO Norm\n6519:2002 Anhang D durch die Einbeziehung einer schnellen und effektiven\nMethode zum Nachweis von Salmonellen direkt nach der Voranreicherung zu verringern.\nEs wurde gezeigt, dass die real-time-PCR eine sehr effektive Methode f\u00fcr\nden Nachweis von Salmonellen aus Lebensmitteln nach einer Voranreicherung\nist. Im Gegensatz dazu sind in Proben aus dem Tierbereich jedoch viel gr\u00f6\u00dfere\nMengen an bakterieller Sekund\u00e4rflora und an hemmenden Substanzen vorhanden,\ndie die Sensitivit\u00e4t der real-time-PCR beeintr\u00e4chtigen k\u00f6nnen.\nDaher wurden verschiedene Methoden zur Isolation von DNA nach einer Voranreicherung\nvon Salmonellen mit Tierkot verglichen, um das Detektionslimit und\ndie Detektionswahrscheinlichkeit mithilfe einer invA<\/strong>-basierten real-time-PCR\nzu bestimmen. Eine Detektionswahrscheinlichkeit von 100 % konnte mit dem\nQIAamp\u00ae DNA Stool Mini Kit (Qiagen, Deutschland) erst ab 105 kbE\/ml, mit dem\nHigh Pure PCR Template Preparation Kit\u00ae (Roche, Deutschland) ab 104 kbE\/ml und\nsowohl bei der Kochlyse als auch bei einem neu entwickelten Hausprotokoll ab\n103 kbE\/ml erreicht werden. Im Vergleich aller verwendeten DNA-Extraktionsmethoden\nbesitzt die Kochlyse eine sehr gute Nachweisgrenze, ein geringes Kontaminationsrisiko\nund verursacht sehr geringe Kosten. Dar\u00fcber hinaus wurden\nProben aus dem Z\u00e4kum-Inhalt von Mastschweinen nach ISO 6519:2002 Annex D\nund der invA<\/strong>-basierten real-time-PCR mit Kochlyse zur DNA-Isolation untersucht.\nDer Vergleich zeigte, dass weder falsch positive noch falsch negative Ergebnisse\nauftraten. Die Einbeziehung der invA<\/strong>-real-time-PCR nach einer Voranreicherung\nder Proben und die nachfolgende Fortsetzung des Nachweises von Salmonellen\nnur mit den Proben, die mittels real-time-PCR positiv waren, k\u00f6nnte ein Verfahren\ndarstellen, um sowohl die Anforderungen der ISO 6519:2002 Anhang D zu erf\u00fcllen\naber gleicherma\u00dfen den Aufwand f\u00fcr diese Methode zu verringen. Es muss\njedoch darauf hingewiesen werden, dass die Modifikation einer international\ng\u00fcltigen Standardmethode und deren Anwendung in der Routinediagnostik eine\nentsprechende Validierung und Zulassung durch die zust\u00e4ndigen nationalen und\ninternationalen Beh\u00f6rden erfordert.\n\n\n\n

\nSchl\u00fcsselw\u00f6rter:<\/strong>
\n\nSalmonella<\/strong>, Polymerasekettenreaktion, DNS-Isolierung, Tierkot,\nISO 6519 Anhang D\n\n\n\n\n\n


\n\n\n\n\n\n\n\n\nQuelle:
Berl. M\u00fcnch. Tier\u00e4rztl. Wschr. 124: 5-6, 177-185 (2011)
\nAutor: Braun SD, Methner U
\nDOI-Nummer: 10.2376\/0005-9366-124-177

\n\u00a9 Schl\u00fctersche Verlagsgesellschaft mbH amp; Co. KG \n\n\n\n\n\n <\/a><\/p>

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<\/p>","primaryLanguage":"englisch","summary":"There is a strong interest to reduce the expenditure for the detection of Salmonella<\/strong>\nspp. from animal faeces and environmental samples from primary production\naccording to ISO 6519:2002 Annex D by including a rapid and effective\nmethod to detect Salmonella<\/strong> spp. already after pre-enrichment in BPW. It has\nbeen shown that real-time PCR methods are very effective to detect Salmonella<\/strong>\norganisms after pre-enrichment of foods. However, materials from primary animal\nproduction compose of much higher amounts of substances which might inhibit\nthe sensitivity of real-time PCR. Different techniques of DNA isolation after preenrichment\nof artificially inoculated bovine faecal material were used to compare\ntheir detection limit and detection probability using an invA<\/strong> 5\u2019 nuclease real-time\nPCR approach. A detection probability of 100% was shown at 105 cfu\/ml using\nthe QIAamp\u00ae DNA Stool Mini Kit (Qiagen, Germany), at 104 cfu\/ml using the High\nPure PCR Template Preparation Kit\u00ae (Roche, Germany) and at 103 cfu\/ml using\nthermal cell lysis or an in-house lab protocol, respectively. In comparison DNA\nisolation by thermal cell lysis revealed a very good detection limit, low costs and\nalmost no risks of contamination. Furthermore, caecal contents from pigs were\nanalysed by ISO 6519:2002 Annex D and the invA<\/strong> real-time PCR using thermal\ncell lysis for DNA extraction. As a result neither false positive nor false negative\nfindings were obtained. Inclusion of the real-time PCR after pre-enrichment of\nsamples in BPW followed by bacterial detection of Salmonella<\/strong> only with samples\npositive with real-time PCR might be a valuable tool to fulfil the international\nstandard of ISO 6519:2002 Annex D but also to diminish the expenditures.\nHowever, it must be stated that the modification of an international standard\nmethod and its use in routine diagnostic requires the validation and registration\nof national and\/ or international competent authorities.","keywords":["Salmonella"],"zusammenfassung":"Es gibt ein starkes Interesse, den Material- und Zeitaufwand f\u00fcr den Nachweis von\nSalmonella<\/strong> spp. aus Tierkot und Umgebungsproben entsprechend der ISO Norm\n6519:2002 Anhang D durch die Einbeziehung einer schnellen und effektiven\nMethode zum Nachweis von Salmonellen direkt nach der Voranreicherung zu verringern.\nEs wurde gezeigt, dass die real-time-PCR eine sehr effektive Methode f\u00fcr\nden Nachweis von Salmonellen aus Lebensmitteln nach einer Voranreicherung\nist. Im Gegensatz dazu sind in Proben aus dem Tierbereich jedoch viel gr\u00f6\u00dfere\nMengen an bakterieller Sekund\u00e4rflora und an hemmenden Substanzen vorhanden,\ndie die Sensitivit\u00e4t der real-time-PCR beeintr\u00e4chtigen k\u00f6nnen.\nDaher wurden verschiedene Methoden zur Isolation von DNA nach einer Voranreicherung\nvon Salmonellen mit Tierkot verglichen, um das Detektionslimit und\ndie Detektionswahrscheinlichkeit mithilfe einer invA<\/strong>-basierten real-time-PCR\nzu bestimmen. Eine Detektionswahrscheinlichkeit von 100 % konnte mit dem\nQIAamp\u00ae DNA Stool Mini Kit (Qiagen, Deutschland) erst ab 105 kbE\/ml, mit dem\nHigh Pure PCR Template Preparation Kit\u00ae (Roche, Deutschland) ab 104 kbE\/ml und\nsowohl bei der Kochlyse als auch bei einem neu entwickelten Hausprotokoll ab\n103 kbE\/ml erreicht werden. Im Vergleich aller verwendeten DNA-Extraktionsmethoden\nbesitzt die Kochlyse eine sehr gute Nachweisgrenze, ein geringes Kontaminationsrisiko\nund verursacht sehr geringe Kosten. Dar\u00fcber hinaus wurden\nProben aus dem Z\u00e4kum-Inhalt von Mastschweinen nach ISO 6519:2002 Annex D\nund der invA<\/strong>-basierten real-time-PCR mit Kochlyse zur DNA-Isolation untersucht.\nDer Vergleich zeigte, dass weder falsch positive noch falsch negative Ergebnisse\nauftraten. Die Einbeziehung der invA<\/strong>-real-time-PCR nach einer Voranreicherung\nder Proben und die nachfolgende Fortsetzung des Nachweises von Salmonellen\nnur mit den Proben, die mittels real-time-PCR positiv waren, k\u00f6nnte ein Verfahren\ndarstellen, um sowohl die Anforderungen der ISO 6519:2002 Anhang D zu erf\u00fcllen\naber gleicherma\u00dfen den Aufwand f\u00fcr diese Methode zu verringen. Es muss\njedoch darauf hingewiesen werden, dass die Modifikation einer international\ng\u00fcltigen Standardmethode und deren Anwendung in der Routinediagnostik eine\nentsprechende Validierung und Zulassung durch die zust\u00e4ndigen nationalen und\ninternationalen Beh\u00f6rden erfordert.","schluesselwoerter":["Salmonella"],"translatedTitle":"","abstractE":"There is a strong interest to reduce the expenditure for the detection of Salmonella\nspp. from animal faeces and environmental samples from primary production\naccording to ISO 6519:2002 Annex D by including a rapid and effective\nmethod to detect Salmonella spp. already after pre-enrichment in BPW. It has\nbeen shown that real-time PCR methods are very effective to detect Salmonella\norganisms after pre-enrichment of foods. However, materials from primary animal\nproduction compose of much higher amounts of substances which might inhibit\nthe sensitivity of real-time PCR. Different techniques of DNA isolation after preenrichment\nof artificially inoculated bovine faecal material were used to compare\ntheir detection limit and detection probability using an invA 5\u2019 nuclease real-time\nPCR approach. A detection probability of 100% was shown at 105 cfu\/ml using\nthe QIAamp\u00ae DNA Stool Mini Kit (Qiagen, Germany), at 104 cfu\/ml using the High\nPure PCR Template Preparation Kit\u00ae (Roche, Germany) and at 103 cfu\/ml using\nthermal cell lysis or an in-house lab protocol, respectively. In comparison DNA\nisolation by thermal cell lysis revealed a very good detection limit, low costs and\nalmost no risks of contamination. Furthermore, caecal contents from pigs were\nanalysed by ISO 6519:2002 Annex D and the invA real-time PCR using thermal\ncell lysis for DNA extraction. As a result neither false positive nor false negative\nfindings were obtained. Inclusion of the real-time PCR after pre-enrichment of\nsamples in BPW followed by bacterial detection of Salmonella only with samples\npositive with real-time PCR might be a valuable tool to fulfil the international\nstandard of ISO 6519:2002 Annex D but also to diminish the expenditures.\nHowever, it must be stated that the modification of an international standard\nmethod and its use in routine diagnostic requires the validation and registration\nof national and\/ or international competent authorities.","date":{"year":2011,"date":"04\/2011","accepted":"2011-04-19"},"volume":"124","openAccess":true,"journal":"Berliner und M\u00fcnchener Tier\u00e4rztliche Wochenschrift","titleImageId":944,"pages":"177-185","redirects":["open-access-salmonellai-polymerase-chain-reaction-dna-isolation-faecal-samples\/150\/3216\/69746","open-access-salmonellai-polymerase-chain-reaction-dna-isolation-faecal-samples\/150\/3130\/69746"],"tierartCategories":[],"artikelartCategories":["Open Access","Tier\u00e4rztliche Wochenschrift","Abostufe BMTW","Fachartikel","Abostufe frei"]} CY - Hannover DA - 04/2011 DO - 10.2376/0005-9366-124-177 LA - English N2 - There is a strong interest to reduce the expenditure for the detection of Salmonella spp. from animal faeces and environmental samples from primary production according to ISO 6519:2002 Annex D by including a rapid and effective method to detect Salmonella spp. already after pre-enrichment in BPW. It has been shown that real-time PCR methods are very effective to detect Salmonella organisms after pre-enrichment of foods. However, materials from primary animal production compose of much higher amounts of substances which might inhibit the sensitivity of real-time PCR. Different techniques of DNA isolation after preenrichment of artificially inoculated bovine faecal material were used to compare their detection limit and detection probability using an invA 5’ nuclease real-time PCR approach. A detection probability of 100% was shown at 105 cfu/ml using the QIAamp® DNA Stool Mini Kit (Qiagen, Germany), at 104 cfu/ml using the High Pure PCR Template Preparation Kit® (Roche, Germany) and at 103 cfu/ml using thermal cell lysis or an in-house lab protocol, respectively. In comparison DNA isolation by thermal cell lysis revealed a very good detection limit, low costs and almost no risks of contamination. Furthermore, caecal contents from pigs were analysed by ISO 6519:2002 Annex D and the invA real-time PCR using thermal cell lysis for DNA extraction. As a result neither false positive nor false negative findings were obtained. Inclusion of the real-time PCR after pre-enrichment of samples in BPW followed by bacterial detection of Salmonella only with samples positive with real-time PCR might be a valuable tool to fulfil the international standard of ISO 6519:2002 Annex D but also to diminish the expenditures. However, it must be stated that the modification of an international standard method and its use in routine diagnostic requires the validation and registration of national and/ or international competent authorities. PB - Schlütersche Verlagsgesellschaft mbH & Co. KG PP - Hannover PY - 2011 SP - 177 EP - 185 T1 - Comparison of DNA isolation methods and detection of Salmonella spp. from animal faeces and dust using invA realtime PCR T2 - Berliner und Münchener Tierärztliche Wochenschrift TI - Comparison of DNA isolation methods and detection of Salmonella spp. from animal faeces and dust using invA realtime PCR VL - 124 SN - 0005-9366 ER -