Comparison of an improved polymerase chain reaction protocol and the indirect hemagglutination assay in the detection of Eperythrozoon suis infection
Department of Veterinary Diagnostic Investigation, College of Veterinary Medicine, Kansas State University, Manhattan 66506. The ability of an improved polymerase chain reaction (PCR) protocol to detect Eperythrozoon suis DNA in the blood of experimentally infected nonsplenectomized pigs was evaluat...
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| Veröffentlicht in: | Journal of veterinary diagnostic investigation 1994-07, Vol.6 (3), p.321-325 |
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| container_title | Journal of veterinary diagnostic investigation |
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| creator | Gwaltney, S.M Oberst, R.D |
| description | Department of Veterinary Diagnostic Investigation, College of Veterinary Medicine, Kansas State University, Manhattan 66506.
The ability of an improved polymerase chain reaction (PCR) protocol to detect Eperythrozoon suis DNA in the blood of experimentally infected nonsplenectomized pigs was evaluated. The protocol utilizes previously described E. suis-specific primers and a proprietary DNA-releasing reagent in a 2-step amplification cycle followed by visualization of the 492-bp amplification product on agarose gels. This PCR protocol successfully amplified E. suis DNA in blood from all postinfection samples and from the preinfection samples of 2 pigs, indicating preexisting natural infections. Results of the indirect hemagglutination test on serum samples from these pigs revealed that only 1 pig developed detectable antibody titers to E. suis infection during the 43-day study; that pig was determined by PCR to have been infected naturally with E. suis prior to experimental inoculation. These results confirm previous reports of poor antibody response of young pigs to E. suis infection and demonstrate the potential of PCR as a valuable tool for the diagnosis and study of E. suis infection in pigs. |
| doi_str_mv | 10.1177/104063879400600307 |
| format | Article |
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The ability of an improved polymerase chain reaction (PCR) protocol to detect Eperythrozoon suis DNA in the blood of experimentally infected nonsplenectomized pigs was evaluated. The protocol utilizes previously described E. suis-specific primers and a proprietary DNA-releasing reagent in a 2-step amplification cycle followed by visualization of the 492-bp amplification product on agarose gels. This PCR protocol successfully amplified E. suis DNA in blood from all postinfection samples and from the preinfection samples of 2 pigs, indicating preexisting natural infections. Results of the indirect hemagglutination test on serum samples from these pigs revealed that only 1 pig developed detectable antibody titers to E. suis infection during the 43-day study; that pig was determined by PCR to have been infected naturally with E. suis prior to experimental inoculation. These results confirm previous reports of poor antibody response of young pigs to E. suis infection and demonstrate the potential of PCR as a valuable tool for the diagnosis and study of E. suis infection in pigs.</description><identifier>ISSN: 1040-6387</identifier><identifier>EISSN: 1943-4936</identifier><identifier>DOI: 10.1177/104063879400600307</identifier><identifier>PMID: 7948201</identifier><language>eng</language><publisher>Los Angeles, CA: J Vet Diagn Invest</publisher><subject>AMPLIFICATION CHAINE POLYMERASE ; Animals ; Base Sequence ; CERDO ; DIAGNOSIS ; DIAGNOSTIC ; DIAGNOSTIC TECHNIQUES ; DIAGNOSTIC VALUE ; DIAGNOSTICO ; DNA, Bacterial - blood ; EPERYTHROZOON ; EPREUVE D'HEMAGGLUTINATION ; EXPERIMENTACION IN VIVO ; EXPERIMENTAL INFECTION ; EXPERIMENTATION IN VIVO ; HAEMAGGLUTINATION TESTS ; HEMAGGLUTINATION TESTS ; Hemagglutination Tests - veterinary ; IN VIVO EXPERIMENTATION ; INFECCION EXPERIMENTAL ; INFECCIONES LATENTES ; INFECTION EXPERIMENTALE ; INFECTION LATENTE ; LATENT INFECTIONS ; Molecular Sequence Data ; Mycoplasma - isolation & purification ; Mycoplasma Infections - blood ; Mycoplasma Infections - microbiology ; Mycoplasma Infections - veterinary ; PIGS ; POLYMERASE CHAIN REACTION ; Polymerase Chain Reaction - methods ; Polymerase Chain Reaction - veterinary ; PORCIN ; PRUEBAS DE HEMAGLUTINACION ; REACCION DE CADENAS DE POLIMERASA ; Sensitivity and Specificity ; SWINE ; Swine Diseases - blood ; Swine Diseases - microbiology</subject><ispartof>Journal of veterinary diagnostic investigation, 1994-07, Vol.6 (3), p.321-325</ispartof><rights>1994 American Association of Veterinary Laboratory Diagnosticians</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c431t-d044c5fe9d46b2f3f85c721a4c880fa7a5184191219760b7225c70be993dcc4c3</citedby><cites>FETCH-LOGICAL-c431t-d044c5fe9d46b2f3f85c721a4c880fa7a5184191219760b7225c70be993dcc4c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7948201$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gwaltney, S.M</creatorcontrib><creatorcontrib>Oberst, R.D</creatorcontrib><title>Comparison of an improved polymerase chain reaction protocol and the indirect hemagglutination assay in the detection of Eperythrozoon suis infection</title><title>Journal of veterinary diagnostic investigation</title><addtitle>J Vet Diagn Invest</addtitle><description>Department of Veterinary Diagnostic Investigation, College of Veterinary Medicine, Kansas State University, Manhattan 66506.
The ability of an improved polymerase chain reaction (PCR) protocol to detect Eperythrozoon suis DNA in the blood of experimentally infected nonsplenectomized pigs was evaluated. The protocol utilizes previously described E. suis-specific primers and a proprietary DNA-releasing reagent in a 2-step amplification cycle followed by visualization of the 492-bp amplification product on agarose gels. This PCR protocol successfully amplified E. suis DNA in blood from all postinfection samples and from the preinfection samples of 2 pigs, indicating preexisting natural infections. Results of the indirect hemagglutination test on serum samples from these pigs revealed that only 1 pig developed detectable antibody titers to E. suis infection during the 43-day study; that pig was determined by PCR to have been infected naturally with E. suis prior to experimental inoculation. These results confirm previous reports of poor antibody response of young pigs to E. suis infection and demonstrate the potential of PCR as a valuable tool for the diagnosis and study of E. suis infection in pigs.</description><subject>AMPLIFICATION CHAINE POLYMERASE</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>CERDO</subject><subject>DIAGNOSIS</subject><subject>DIAGNOSTIC</subject><subject>DIAGNOSTIC TECHNIQUES</subject><subject>DIAGNOSTIC VALUE</subject><subject>DIAGNOSTICO</subject><subject>DNA, Bacterial - blood</subject><subject>EPERYTHROZOON</subject><subject>EPREUVE D'HEMAGGLUTINATION</subject><subject>EXPERIMENTACION IN VIVO</subject><subject>EXPERIMENTAL INFECTION</subject><subject>EXPERIMENTATION IN VIVO</subject><subject>HAEMAGGLUTINATION TESTS</subject><subject>HEMAGGLUTINATION TESTS</subject><subject>Hemagglutination Tests - veterinary</subject><subject>IN VIVO EXPERIMENTATION</subject><subject>INFECCION EXPERIMENTAL</subject><subject>INFECCIONES LATENTES</subject><subject>INFECTION EXPERIMENTALE</subject><subject>INFECTION LATENTE</subject><subject>LATENT INFECTIONS</subject><subject>Molecular Sequence Data</subject><subject>Mycoplasma - isolation & purification</subject><subject>Mycoplasma Infections - blood</subject><subject>Mycoplasma Infections - microbiology</subject><subject>Mycoplasma Infections - veterinary</subject><subject>PIGS</subject><subject>POLYMERASE CHAIN REACTION</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Polymerase Chain Reaction - veterinary</subject><subject>PORCIN</subject><subject>PRUEBAS DE HEMAGLUTINACION</subject><subject>REACCION DE CADENAS DE POLIMERASA</subject><subject>Sensitivity and Specificity</subject><subject>SWINE</subject><subject>Swine Diseases - blood</subject><subject>Swine Diseases - microbiology</subject><issn>1040-6387</issn><issn>1943-4936</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc1u1DAUhSMEKqXwApWQsoFdqP9ix0s0Kj9SJRbQtXXHdhKPkjjYSavhPXhf7jQjNkisbPl851j3nqK4puQDpUrdUCKI5I3SghBJCCfqWXFJteCV0Fw-xzsC1Yl4WbzK-UBIzWpFL4oLtDSM0Mvi9y6OM6SQ41TGtoSpDOOc4oN35RyH4-gTZF_aHsJUJg92CQgisEQbB8RdufS-DJMLydul7P0IXTesS5jgCYWc4Yj6E-b84rcE_Op29um49Cn-iviQ15ARazf9dfGihSH7N-fzqrj_dPtj96W6-_b56-7jXWUFp0vliBC2br12Qu5Zy9umtopRELZpSAsKatoIqimjWkmyV4yhTvZea-6sFZZfFe-3XJzo5-rzYsaQrR8GmHxcs1FSaSnqBkG2gTbFnJNvzZzCCOloKDGnLsy_XaDp7Tl93Y_e_bWcl4_6zaZn6Lw5xDVNOOz_E99tjj50_SOu3OQRhgHzmTk8uCANN5ydkq83roVooMN6zf13XROmpOR_AAioqj0</recordid><startdate>19940701</startdate><enddate>19940701</enddate><creator>Gwaltney, S.M</creator><creator>Oberst, R.D</creator><general>J Vet Diagn Invest</general><general>SAGE Publications</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19940701</creationdate><title>Comparison of an improved polymerase chain reaction protocol and the indirect hemagglutination assay in the detection of Eperythrozoon suis infection</title><author>Gwaltney, S.M ; Oberst, R.D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c431t-d044c5fe9d46b2f3f85c721a4c880fa7a5184191219760b7225c70be993dcc4c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>AMPLIFICATION CHAINE POLYMERASE</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>CERDO</topic><topic>DIAGNOSIS</topic><topic>DIAGNOSTIC</topic><topic>DIAGNOSTIC TECHNIQUES</topic><topic>DIAGNOSTIC VALUE</topic><topic>DIAGNOSTICO</topic><topic>DNA, Bacterial - blood</topic><topic>EPERYTHROZOON</topic><topic>EPREUVE D'HEMAGGLUTINATION</topic><topic>EXPERIMENTACION IN VIVO</topic><topic>EXPERIMENTAL INFECTION</topic><topic>EXPERIMENTATION IN VIVO</topic><topic>HAEMAGGLUTINATION TESTS</topic><topic>HEMAGGLUTINATION TESTS</topic><topic>Hemagglutination Tests - veterinary</topic><topic>IN VIVO EXPERIMENTATION</topic><topic>INFECCION EXPERIMENTAL</topic><topic>INFECCIONES LATENTES</topic><topic>INFECTION EXPERIMENTALE</topic><topic>INFECTION LATENTE</topic><topic>LATENT INFECTIONS</topic><topic>Molecular Sequence Data</topic><topic>Mycoplasma - isolation & purification</topic><topic>Mycoplasma Infections - blood</topic><topic>Mycoplasma Infections - microbiology</topic><topic>Mycoplasma Infections - veterinary</topic><topic>PIGS</topic><topic>POLYMERASE CHAIN REACTION</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Polymerase Chain Reaction - veterinary</topic><topic>PORCIN</topic><topic>PRUEBAS DE HEMAGLUTINACION</topic><topic>REACCION DE CADENAS DE POLIMERASA</topic><topic>Sensitivity and Specificity</topic><topic>SWINE</topic><topic>Swine Diseases - blood</topic><topic>Swine Diseases - microbiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gwaltney, S.M</creatorcontrib><creatorcontrib>Oberst, R.D</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of veterinary diagnostic investigation</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gwaltney, S.M</au><au>Oberst, R.D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comparison of an improved polymerase chain reaction protocol and the indirect hemagglutination assay in the detection of Eperythrozoon suis infection</atitle><jtitle>Journal of veterinary diagnostic investigation</jtitle><addtitle>J Vet Diagn Invest</addtitle><date>1994-07-01</date><risdate>1994</risdate><volume>6</volume><issue>3</issue><spage>321</spage><epage>325</epage><pages>321-325</pages><issn>1040-6387</issn><eissn>1943-4936</eissn><abstract>Department of Veterinary Diagnostic Investigation, College of Veterinary Medicine, Kansas State University, Manhattan 66506.
The ability of an improved polymerase chain reaction (PCR) protocol to detect Eperythrozoon suis DNA in the blood of experimentally infected nonsplenectomized pigs was evaluated. The protocol utilizes previously described E. suis-specific primers and a proprietary DNA-releasing reagent in a 2-step amplification cycle followed by visualization of the 492-bp amplification product on agarose gels. This PCR protocol successfully amplified E. suis DNA in blood from all postinfection samples and from the preinfection samples of 2 pigs, indicating preexisting natural infections. Results of the indirect hemagglutination test on serum samples from these pigs revealed that only 1 pig developed detectable antibody titers to E. suis infection during the 43-day study; that pig was determined by PCR to have been infected naturally with E. suis prior to experimental inoculation. These results confirm previous reports of poor antibody response of young pigs to E. suis infection and demonstrate the potential of PCR as a valuable tool for the diagnosis and study of E. suis infection in pigs.</abstract><cop>Los Angeles, CA</cop><pub>J Vet Diagn Invest</pub><pmid>7948201</pmid><doi>10.1177/104063879400600307</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
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| ispartof | Journal of veterinary diagnostic investigation, 1994-07, Vol.6 (3), p.321-325 |
| issn | 1040-6387 1943-4936 |
| language | eng |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | AMPLIFICATION CHAINE POLYMERASE Animals Base Sequence CERDO DIAGNOSIS DIAGNOSTIC DIAGNOSTIC TECHNIQUES DIAGNOSTIC VALUE DIAGNOSTICO DNA, Bacterial - blood EPERYTHROZOON EPREUVE D'HEMAGGLUTINATION EXPERIMENTACION IN VIVO EXPERIMENTAL INFECTION EXPERIMENTATION IN VIVO HAEMAGGLUTINATION TESTS HEMAGGLUTINATION TESTS Hemagglutination Tests - veterinary IN VIVO EXPERIMENTATION INFECCION EXPERIMENTAL INFECCIONES LATENTES INFECTION EXPERIMENTALE INFECTION LATENTE LATENT INFECTIONS Molecular Sequence Data Mycoplasma - isolation & purification Mycoplasma Infections - blood Mycoplasma Infections - microbiology Mycoplasma Infections - veterinary PIGS POLYMERASE CHAIN REACTION Polymerase Chain Reaction - methods Polymerase Chain Reaction - veterinary PORCIN PRUEBAS DE HEMAGLUTINACION REACCION DE CADENAS DE POLIMERASA Sensitivity and Specificity SWINE Swine Diseases - blood Swine Diseases - microbiology |
| title | Comparison of an improved polymerase chain reaction protocol and the indirect hemagglutination assay in the detection of Eperythrozoon suis infection |
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