TY - JOUR KW - RT-qPCR KW - qPCR KW - pig KW - pen-wise KW - individual AU - A Steinrigl AU - S Revilla-Fernández AU - F Schmoll AU - T Sattler AB - The earliest possible detection of pathogens is of utmost importance in limiting the spread of contagious diseases, such as Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Circovirus Type 2 (PCV-2). Alternative sample matrices must enable sensitive detection of these swine pathogens without the need for standard blood collection in pigs by venipuncture. Here, we investigate the potential use of GenoTube swabs for PCR detection of PRRSV and PCV-2 in individual blood and oral fluid swabs from pigs after experimental PRRSV infection. The results were compared with matched serum and pen-wise collected oral fluid. It was shown that PCV-2 detection in GenoTube blood swabs, oral fluid swabs and pen-wise oral fluid is at least as sensitive as in serum, while PRRSV detection is most sensitive in serum, followed by the blood swabs and oral fluids. A good correlation was observed between PCV-2 and PRRSV loads in serum and blood swabs, while there was little or no correlation between serum and oral fluids. A modified extraction protocol enabled further improvement of PRRSV RNA recovery from GenoTube swabs. Furthermore, we show that GenoTube swabs are suitable for long-term (up to 56 days) storage of specimens at room temperature without significant influence on PRRSV RNA stability. In conclusion, blood and oral fluid collected by GenoTube swabs are practical and reasonable sample matrices for PRRSV and PCV-2 detection by RTq-PCR and qPCR, respectively. However, it has to be kept in mind that viral loads in oral fluid do not accurately reflect those in serum. BT - Berliner und Münchener Tierärztliche Wochenschrift C1 - {"oldId":95552,"title":"Detection of Porcine Reproductive and Respiratory Syndrome Virus and Porcine Cirovirus Type 2 in blood and oral fluid collected with GenoTube swabs","topline":"Open Access","teaserText":"Nachweis des Porzinen Reproduktiven und Respiratorischen Syndrom Virus und des Porzinen Circovirus Typ 2 in mittels GenoTube-Tupfern gewonnenem Blut und Speichel","content":"

Summary<\/span>
The earliest possible detection of pathogens is of utmost importance in limiting the spread of contagious diseases, such as Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Circovirus Type 2 (PCV-2). Alternative sample matrices must enable sensitive detection of these swine pathogens without the need for standard blood collection in pigs by venipuncture. Here, we investigate the potential use of GenoTube swabs for PCR detection of PRRSV and PCV-2 in individual blood and oral fluid swabs from pigs after experimental PRRSV infection. The results were compared with matched serum and pen-wise collected oral fluid. It was shown that PCV-2 detection in GenoTube blood swabs, oral fluid swabs and pen-wise oral fluid is at least as sensitive as in serum, while PRRSV detection is most sensitive in serum, followed by the blood swabs and oral fluids. A good correlation was observed between PCV-2 and PRRSV loads in serum and blood swabs, while there was little or no correlation between serum and oral fluids. A modified extraction protocol enabled further improvement of PRRSV RNA recovery from GenoTube swabs. Furthermore, we show that GenoTube swabs are suitable for long-term (up to 56 days) storage of specimens at room temperature without significant influence on PRRSV RNA stability. In conclusion, blood and oral fluid collected by GenoTube swabs are practical and reasonable sample matrices for PRRSV and PCV-2 detection by RTq-PCR and qPCR, respectively. However, it has to be kept in mind that viral loads in oral fluid do not accurately reflect those in serum.

Keywords<\/span>: RT-qPCR, qPCR, pig, pen-wise, individual<\/p>

Zusammenfassung<\/span>
Eine m\u00f6glichst fr\u00fchzeitige Detektion von Pathogenen ist von gr\u00f6\u00dfter Wichtigkeit f\u00fcr die Eind\u00e4mmung der Ausbreitung von Infektionskrankheiten wie das Porzine Reproduktive und Respiratorische Syndrom Virus (PRRSV) und das Porzine Circovirus Typ 2 (PCV-2). Alternative Probenmaterialien m\u00fcssen f\u00fcr den sensitiven Nachweis dieser Erreger geeignet sein und sollten ohne die herk\u00f6mmliche und aufwendige Blutentnahme bei Schweinen auskommen. Hier untersuchen wir die m\u00f6gliche Verwendung von GenoTube-Trockentupfern f\u00fcr den PCR-Nachweis von PRRSV und PCV-2 in individuellen Blut- und Speichelproben von Schweinen nach experimenteller PRRSV-Infektion. Die Ergebnisse wurden mit den korrespondierenden Serumproben und buchtenweise gewonnener Speichelfl\u00fcssigkeit verglichen. Es zeigte sich, dass der PCV-2-Nachweis in Bluttupfern, Speicheltupfern und buchtenweiser Speichelfl\u00fcssigkeit mindestens genauso sensitiv ist wie im Serum, w\u00e4hrend PRRSV am sensitivsten in Serum nachgewiesen wurde, gefolgt von Bluttupfern und Speichelfl\u00fcssigkeit. Es bestand eine enge Korrelation zwischen den PCV-2- und PRRSV-Lasten in Serum und Bluttupfern, wohingegen die Korrelation zwischen Serum und Speichelfl\u00fcssigkeit geringer oder nicht vorhanden war. Ein modifiziertes Extraktionsprotokoll f\u00fchrte zu zus\u00e4tzlicher Verbesserung der PRRSV-RNA-R\u00fcckgewinnung aus den GenoTube-Tupfern. Weiterhin zeigen wir, dass GenoTube-Tupfer auch f\u00fcr die Langzeitlagerung (bis zu 56 Tage) bei Raumtemperatur geeignet sind, ohne dass es zu signifikanten Einfl\u00fcssen auf die PRRSV-RNA-Stabilit\u00e4t kommt. Zusammenfassend kann gesagt werden, dass mit dem GenoTube-System gewonnene Blut- und Speicheltupfer eine praktische und sinnvolle Probenmatrix f\u00fcr den PRRSV- und PCV-2-Nachweis mittels RT-qPCR bzw. qPCR sind. Es sollte jedoch daran gedacht werden, dass die Viruslasten in der Speichelfl\u00fcssigkeit nicht unbedingt jenen im Serum entsprechen.

Schl\u00fcsselw\u00f6rter: <\/span>RT-qPCR, qPCR, Schwein, gruppenweise, individuell<\/p>","categories":["Open Access","Tier\u00e4rztliche Wochenschrift","Abostufe BMTW","Fachartikel","Abostufe frei"],"fromDate":"Jun 1, 2016 7:54:59 AM","oldUrls":["http:\/\/vetline.de\/detection-of-porcine-reproductive-and-respiratory-syndrome-virus-and-porcine-cirovirus-type-2-in-blood-and-oral-fluid-collected-with-genotube-swabs\/150\/3130\/95552","http:\/\/vetline.de\/detection-of-porcine-reproductive-and-respiratory-syndrome-virus-and-porcine-cirovirus-type-2-in-blood-and-oral-fluid-collected-with-genotube-swabs\/150\/3216\/95552"],"doiLanguage":"englisch","doiProductFormat":"online","doiPublisher":"Schl\u00fctersche Verlagsgesellschaft mbH & Co. KG","doiSerialWorkTitle":"Berl M\u00fcnch Tier\u00e4rztl Wochensch","doiDocumentUri":"http:\/\/cf01.vetline.schluetersche.de\/files\/smfiledata\/5\/7\/6\/7\/1\/9\/BMW_2016_09_10_0437_onl300.pdf","doiSource":"Berl M\u00fcnch Tier\u00e4rztl Wochenschr 129, 437-443","doiissn":"0005-9366","doiNr":"10.2376\/0005-9366-16029","doiFirstPage":"437","doiLastPage":"443","doiTransmitted":true,"doiAuthor":"Steinrigl A, Revilla-Fern\u00e1ndez S, Schmoll F, Sattler T","pdf":{"path":"http:\/\/data\/BMW_2016_09_10_0437_onl300.pdf","title":"BMTW OA 16029 Sattler","description":""},"authors":[{"firstName":"A","middleName":"","lastName":"Steinrigl"},{"firstName":"S","middleName":"","lastName":"Revilla-Fern\u00e1ndez"},{"firstName":"F","middleName":"","lastName":"Schmoll"},{"firstName":"T","middleName":"","lastName":"Sattler"}],"contentOptimised":"

Summary<\/strong>
The earliest possible detection of pathogens is of utmost importance in limiting the spread of contagious diseases, such as Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Circovirus Type 2 (PCV-2). Alternative sample matrices must enable sensitive detection of these swine pathogens without the need for standard blood collection in pigs by venipuncture. Here, we investigate the potential use of GenoTube swabs for PCR detection of PRRSV and PCV-2 in individual blood and oral fluid swabs from pigs after experimental PRRSV infection. The results were compared with matched serum and pen-wise collected oral fluid. It was shown that PCV-2 detection in GenoTube blood swabs, oral fluid swabs and pen-wise oral fluid is at least as sensitive as in serum, while PRRSV detection is most sensitive in serum, followed by the blood swabs and oral fluids. A good correlation was observed between PCV-2 and PRRSV loads in serum and blood swabs, while there was little or no correlation between serum and oral fluids. A modified extraction protocol enabled further improvement of PRRSV RNA recovery from GenoTube swabs. Furthermore, we show that GenoTube swabs are suitable for long-term (up to 56 days) storage of specimens at room temperature without significant influence on PRRSV RNA stability. In conclusion, blood and oral fluid collected by GenoTube swabs are practical and reasonable sample matrices for PRRSV and PCV-2 detection by RTq-PCR and qPCR, respectively. However, it has to be kept in mind that viral loads in oral fluid do not accurately reflect those in serum.

Keywords:<\/strong>: RT-qPCR, qPCR, pig, pen-wise, individual<\/p>

Zusammenfassung<\/strong>
Eine m\u00f6glichst fr\u00fchzeitige Detektion von Pathogenen ist von gr\u00f6\u00dfter Wichtigkeit f\u00fcr die Eind\u00e4mmung der Ausbreitung von Infektionskrankheiten wie das Porzine Reproduktive und Respiratorische Syndrom Virus (PRRSV) und das Porzine Circovirus Typ 2 (PCV-2). Alternative Probenmaterialien m\u00fcssen f\u00fcr den sensitiven Nachweis dieser Erreger geeignet sein und sollten ohne die herk\u00f6mmliche und aufwendige Blutentnahme bei Schweinen auskommen. Hier untersuchen wir die m\u00f6gliche Verwendung von GenoTube-Trockentupfern f\u00fcr den PCR-Nachweis von PRRSV und PCV-2 in individuellen Blut- und Speichelproben von Schweinen nach experimenteller PRRSV-Infektion. Die Ergebnisse wurden mit den korrespondierenden Serumproben und buchtenweise gewonnener Speichelfl\u00fcssigkeit verglichen. Es zeigte sich, dass der PCV-2-Nachweis in Bluttupfern, Speicheltupfern und buchtenweiser Speichelfl\u00fcssigkeit mindestens genauso sensitiv ist wie im Serum, w\u00e4hrend PRRSV am sensitivsten in Serum nachgewiesen wurde, gefolgt von Bluttupfern und Speichelfl\u00fcssigkeit. Es bestand eine enge Korrelation zwischen den PCV-2- und PRRSV-Lasten in Serum und Bluttupfern, wohingegen die Korrelation zwischen Serum und Speichelfl\u00fcssigkeit geringer oder nicht vorhanden war. Ein modifiziertes Extraktionsprotokoll f\u00fchrte zu zus\u00e4tzlicher Verbesserung der PRRSV-RNA-R\u00fcckgewinnung aus den GenoTube-Tupfern. Weiterhin zeigen wir, dass GenoTube-Tupfer auch f\u00fcr die Langzeitlagerung (bis zu 56 Tage) bei Raumtemperatur geeignet sind, ohne dass es zu signifikanten Einfl\u00fcssen auf die PRRSV-RNA-Stabilit\u00e4t kommt. Zusammenfassend kann gesagt werden, dass mit dem GenoTube-System gewonnene Blut- und Speicheltupfer eine praktische und sinnvolle Probenmatrix f\u00fcr den PRRSV- und PCV-2-Nachweis mittels RT-qPCR bzw. qPCR sind. Es sollte jedoch daran gedacht werden, dass die Viruslasten in der Speichelfl\u00fcssigkeit nicht unbedingt jenen im Serum entsprechen.

Schl\u00fcsselw\u00f6rter:<\/strong>RT-qPCR, qPCR, Schwein, gruppenweise, individuell<\/p>","primaryLanguage":"englisch","summary":"The earliest possible detection of pathogens is of utmost importance in limiting the spread of contagious diseases, such as Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Circovirus Type 2 (PCV-2). Alternative sample matrices must enable sensitive detection of these swine pathogens without the need for standard blood collection in pigs by venipuncture. Here, we investigate the potential use of GenoTube swabs for PCR detection of PRRSV and PCV-2 in individual blood and oral fluid swabs from pigs after experimental PRRSV infection. The results were compared with matched serum and pen-wise collected oral fluid. It was shown that PCV-2 detection in GenoTube blood swabs, oral fluid swabs and pen-wise oral fluid is at least as sensitive as in serum, while PRRSV detection is most sensitive in serum, followed by the blood swabs and oral fluids. A good correlation was observed between PCV-2 and PRRSV loads in serum and blood swabs, while there was little or no correlation between serum and oral fluids. A modified extraction protocol enabled further improvement of PRRSV RNA recovery from GenoTube swabs. Furthermore, we show that GenoTube swabs are suitable for long-term (up to 56 days) storage of specimens at room temperature without significant influence on PRRSV RNA stability. In conclusion, blood and oral fluid collected by GenoTube swabs are practical and reasonable sample matrices for PRRSV and PCV-2 detection by RTq-PCR and qPCR, respectively. However, it has to be kept in mind that viral loads in oral fluid do not accurately reflect those in serum.","keywords":[": RT-qPCR","qPCR","pig","pen-wise","individual"],"zusammenfassung":"Eine m\u00f6glichst fr\u00fchzeitige Detektion von Pathogenen ist von gr\u00f6\u00dfter Wichtigkeit f\u00fcr die Eind\u00e4mmung der Ausbreitung von Infektionskrankheiten wie das Porzine Reproduktive und Respiratorische Syndrom Virus (PRRSV) und das Porzine Circovirus Typ 2 (PCV-2). Alternative Probenmaterialien m\u00fcssen f\u00fcr den sensitiven Nachweis dieser Erreger geeignet sein und sollten ohne die herk\u00f6mmliche und aufwendige Blutentnahme bei Schweinen auskommen. Hier untersuchen wir die m\u00f6gliche Verwendung von GenoTube-Trockentupfern f\u00fcr den PCR-Nachweis von PRRSV und PCV-2 in individuellen Blut- und Speichelproben von Schweinen nach experimenteller PRRSV-Infektion. Die Ergebnisse wurden mit den korrespondierenden Serumproben und buchtenweise gewonnener Speichelfl\u00fcssigkeit verglichen. Es zeigte sich, dass der PCV-2-Nachweis in Bluttupfern, Speicheltupfern und buchtenweiser Speichelfl\u00fcssigkeit mindestens genauso sensitiv ist wie im Serum, w\u00e4hrend PRRSV am sensitivsten in Serum nachgewiesen wurde, gefolgt von Bluttupfern und Speichelfl\u00fcssigkeit. Es bestand eine enge Korrelation zwischen den PCV-2- und PRRSV-Lasten in Serum und Bluttupfern, wohingegen die Korrelation zwischen Serum und Speichelfl\u00fcssigkeit geringer oder nicht vorhanden war. Ein modifiziertes Extraktionsprotokoll f\u00fchrte zu zus\u00e4tzlicher Verbesserung der PRRSV-RNA-R\u00fcckgewinnung aus den GenoTube-Tupfern. Weiterhin zeigen wir, dass GenoTube-Tupfer auch f\u00fcr die Langzeitlagerung (bis zu 56 Tage) bei Raumtemperatur geeignet sind, ohne dass es zu signifikanten Einfl\u00fcssen auf die PRRSV-RNA-Stabilit\u00e4t kommt. Zusammenfassend kann gesagt werden, dass mit dem GenoTube-System gewonnene Blut- und Speicheltupfer eine praktische und sinnvolle Probenmatrix f\u00fcr den PRRSV- und PCV-2-Nachweis mittels RT-qPCR bzw. qPCR sind. Es sollte jedoch daran gedacht werden, dass die Viruslasten in der Speichelfl\u00fcssigkeit nicht unbedingt jenen im Serum entsprechen.","schluesselwoerter":["RT-qPCR","qPCR","Schwein","gruppenweise","individuell"],"translatedTitle":"Nachweis des Porzinen Reproduktiven und Respiratorischen Syndrom Virus und des Porzinen Circovirus Typ 2 in mittels GenoTube-Tupfern gewonnenem Blut und Speichel","abstractE":"The earliest possible detection of pathogens is of utmost importance in limiting the spread of contagious diseases, such as Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Circovirus Type 2 (PCV-2). Alternative sample matrices must enable sensitive detection of these swine pathogens without the need for standard blood collection in pigs by venipuncture. Here, we investigate the potential use of GenoTube swabs for PCR detection of PRRSV and PCV-2 in individual blood and oral fluid swabs from pigs after experimental PRRSV infection. The results were compared with matched serum and pen-wise collected oral fluid. It was shown that PCV-2 detection in GenoTube blood swabs, oral fluid swabs and pen-wise oral fluid is at least as sensitive as in serum, while PRRSV detection is most sensitive in serum, followed by the blood swabs and oral fluids. A good correlation was observed between PCV-2 and PRRSV loads in serum and blood swabs, while there was little or no correlation between serum and oral fluids. A modified extraction protocol enabled further improvement of PRRSV RNA recovery from GenoTube swabs. Furthermore, we show that GenoTube swabs are suitable for long-term (up to 56 days) storage of specimens at room temperature without significant influence on PRRSV RNA stability. In conclusion, blood and oral fluid collected by GenoTube swabs are practical and reasonable sample matrices for PRRSV and PCV-2 detection by RTq-PCR and qPCR, respectively. However, it has to be kept in mind that viral loads in oral fluid do not accurately reflect those in serum.","date":{"year":2016,"date":"06\/2016","accepted":"2016-06-01"},"volume":"129","openAccess":true,"journal":"Berliner und M\u00fcnchener Tier\u00e4rztliche Wochenschrift","titleImageId":944,"pages":"437-443","redirects":["detection-of-porcine-reproductive-and-respiratory-syndrome-virus-and-porcine-cirovirus-type-2-in-blood-and-oral-fluid-collected-with-genotube-swabs\/150\/3130\/95552","detection-of-porcine-reproductive-and-respiratory-syndrome-virus-and-porcine-cirovirus-type-2-in-blood-and-oral-fluid-collected-with-genotube-swabs\/150\/3216\/95552"],"tierartCategories":[],"artikelartCategories":["Open Access","Tier\u00e4rztliche Wochenschrift","Abostufe BMTW","Fachartikel","Abostufe frei"]} CY - Hannover DA - 06/2016 DO - 10.2376/0005-9366-16029 ET - 9/10 LA - English N2 - The earliest possible detection of pathogens is of utmost importance in limiting the spread of contagious diseases, such as Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Circovirus Type 2 (PCV-2). Alternative sample matrices must enable sensitive detection of these swine pathogens without the need for standard blood collection in pigs by venipuncture. Here, we investigate the potential use of GenoTube swabs for PCR detection of PRRSV and PCV-2 in individual blood and oral fluid swabs from pigs after experimental PRRSV infection. The results were compared with matched serum and pen-wise collected oral fluid. It was shown that PCV-2 detection in GenoTube blood swabs, oral fluid swabs and pen-wise oral fluid is at least as sensitive as in serum, while PRRSV detection is most sensitive in serum, followed by the blood swabs and oral fluids. A good correlation was observed between PCV-2 and PRRSV loads in serum and blood swabs, while there was little or no correlation between serum and oral fluids. A modified extraction protocol enabled further improvement of PRRSV RNA recovery from GenoTube swabs. Furthermore, we show that GenoTube swabs are suitable for long-term (up to 56 days) storage of specimens at room temperature without significant influence on PRRSV RNA stability. In conclusion, blood and oral fluid collected by GenoTube swabs are practical and reasonable sample matrices for PRRSV and PCV-2 detection by RTq-PCR and qPCR, respectively. However, it has to be kept in mind that viral loads in oral fluid do not accurately reflect those in serum. PB - Schlütersche Verlagsgesellschaft mbH & Co. KG PP - Hannover PY - 2016 SP - 437 EP - 443 T1 - Detection of Porcine Reproductive and Respiratory Syndrome Virus and Porcine Cirovirus Type 2 in blood and oral fluid collected with GenoTube swabs T2 - Berliner und Münchener Tierärztliche Wochenschrift TI - Detection of Porcine Reproductive and Respiratory Syndrome Virus and Porcine Cirovirus Type 2 in blood and oral fluid collected with GenoTube swabs TT - Nachweis des Porzinen Reproduktiven und Respiratorischen Syndrom Virus und des Porzinen Circovirus Typ 2 in mittels GenoTube-Tupfern gewonnenem Blut und Speichel VL - 129 SN - 0005-9366 ER -