Cargando…

A DNA miniarray system for simultaneous visual detection of porcine circovirus type 1 (PCV1) and 2 (PCV2) in pigs

A miniarray system was developed for the simultaneous detection of porcine circovirus type 1 (PCV1) and type 2 (PCV2) in pigs. The system consists of a polymerase chain reaction (PCR) step to amplify target viral DNA, followed by detection of the amplified DNA using a membrane-anchored probe array a...

Descripción completa

Detalles Bibliográficos
Autores principales: An, D. J., Song, D. S., Park, J. Y., Park, B. K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7088660/
https://www.ncbi.nlm.nih.gov/pubmed/18651234
http://dx.doi.org/10.1007/s11259-008-9080-8
_version_ 1783509583967289344
author An, D. J.
Song, D. S.
Park, J. Y.
Park, B. K.
author_facet An, D. J.
Song, D. S.
Park, J. Y.
Park, B. K.
author_sort An, D. J.
collection PubMed
description A miniarray system was developed for the simultaneous detection of porcine circovirus type 1 (PCV1) and type 2 (PCV2) in pigs. The system consists of a polymerase chain reaction (PCR) step to amplify target viral DNA, followed by detection of the amplified DNA using a membrane-anchored probe array and an avidin-alkaline phosphatase (Av-AP) indicator system. The lower limit of detection of PCV using the miniarray was 10(1.9) tissue culture infectious dose 50 (TCID(50))/ml and 10(2.08)TCID(50)/ml for PCV1 and PCV2, respectively, and 100 viral copies/μl for both PCV1 and PCV2. We validated the miniarray system using 141 lymph node specimens from pigs with suspected postweaning multisystemic wasting syndrome or porcine dermatitis and nephropathy syndrome. Of the 141 samples evaluated, 55 were identified as positive for PCV by the miniarray. Relative to in situ hybridization, the sensitivity and specificity of the miniarray was 100% and 98.9%, respectively. In contrast to other microarray systems, the miniarray does not require a DNA chip reader, since the results can be determined by visual inspection of colorized spots on a nylon membrane. This system represents an effective alternative method for the differential detection of PCV1 and PCV2 in pigs, as well as the maintenance of PCV-free cell lines and pre-screening of commercial vaccines for possible contamination.
format Online
Article
Text
id pubmed-7088660
institution National Center for Biotechnology Information
language English
publishDate 2008
publisher Springer Netherlands
record_format MEDLINE/PubMed
spelling pubmed-70886602020-03-23 A DNA miniarray system for simultaneous visual detection of porcine circovirus type 1 (PCV1) and 2 (PCV2) in pigs An, D. J. Song, D. S. Park, J. Y. Park, B. K. Vet Res Commun Original Article A miniarray system was developed for the simultaneous detection of porcine circovirus type 1 (PCV1) and type 2 (PCV2) in pigs. The system consists of a polymerase chain reaction (PCR) step to amplify target viral DNA, followed by detection of the amplified DNA using a membrane-anchored probe array and an avidin-alkaline phosphatase (Av-AP) indicator system. The lower limit of detection of PCV using the miniarray was 10(1.9) tissue culture infectious dose 50 (TCID(50))/ml and 10(2.08)TCID(50)/ml for PCV1 and PCV2, respectively, and 100 viral copies/μl for both PCV1 and PCV2. We validated the miniarray system using 141 lymph node specimens from pigs with suspected postweaning multisystemic wasting syndrome or porcine dermatitis and nephropathy syndrome. Of the 141 samples evaluated, 55 were identified as positive for PCV by the miniarray. Relative to in situ hybridization, the sensitivity and specificity of the miniarray was 100% and 98.9%, respectively. In contrast to other microarray systems, the miniarray does not require a DNA chip reader, since the results can be determined by visual inspection of colorized spots on a nylon membrane. This system represents an effective alternative method for the differential detection of PCV1 and PCV2 in pigs, as well as the maintenance of PCV-free cell lines and pre-screening of commercial vaccines for possible contamination. Springer Netherlands 2008-07-24 2009 /pmc/articles/PMC7088660/ /pubmed/18651234 http://dx.doi.org/10.1007/s11259-008-9080-8 Text en © Springer Science+Business Media B.V. 2008 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Original Article
An, D. J.
Song, D. S.
Park, J. Y.
Park, B. K.
A DNA miniarray system for simultaneous visual detection of porcine circovirus type 1 (PCV1) and 2 (PCV2) in pigs
title A DNA miniarray system for simultaneous visual detection of porcine circovirus type 1 (PCV1) and 2 (PCV2) in pigs
title_full A DNA miniarray system for simultaneous visual detection of porcine circovirus type 1 (PCV1) and 2 (PCV2) in pigs
title_fullStr A DNA miniarray system for simultaneous visual detection of porcine circovirus type 1 (PCV1) and 2 (PCV2) in pigs
title_full_unstemmed A DNA miniarray system for simultaneous visual detection of porcine circovirus type 1 (PCV1) and 2 (PCV2) in pigs
title_short A DNA miniarray system for simultaneous visual detection of porcine circovirus type 1 (PCV1) and 2 (PCV2) in pigs
title_sort dna miniarray system for simultaneous visual detection of porcine circovirus type 1 (pcv1) and 2 (pcv2) in pigs
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7088660/
https://www.ncbi.nlm.nih.gov/pubmed/18651234
http://dx.doi.org/10.1007/s11259-008-9080-8
work_keys_str_mv AT andj adnaminiarraysystemforsimultaneousvisualdetectionofporcinecircovirustype1pcv1and2pcv2inpigs
AT songds adnaminiarraysystemforsimultaneousvisualdetectionofporcinecircovirustype1pcv1and2pcv2inpigs
AT parkjy adnaminiarraysystemforsimultaneousvisualdetectionofporcinecircovirustype1pcv1and2pcv2inpigs
AT parkbk adnaminiarraysystemforsimultaneousvisualdetectionofporcinecircovirustype1pcv1and2pcv2inpigs
AT andj dnaminiarraysystemforsimultaneousvisualdetectionofporcinecircovirustype1pcv1and2pcv2inpigs
AT songds dnaminiarraysystemforsimultaneousvisualdetectionofporcinecircovirustype1pcv1and2pcv2inpigs
AT parkjy dnaminiarraysystemforsimultaneousvisualdetectionofporcinecircovirustype1pcv1and2pcv2inpigs
AT parkbk dnaminiarraysystemforsimultaneousvisualdetectionofporcinecircovirustype1pcv1and2pcv2inpigs