Use of FTA technology for detection of Trichomonas gallinae

•In this study, we evaluated non-indicating FTA Elute micro cards as a potential sampling substrate for the molecular identification of Trichomonas gallinae isolates.•Isolate concentrations of 10 or 100 trichomonads/40μl, from two T. gallinae isolates were inoculated onto a FTA Elute cards and analy...

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Veröffentlicht in:Veterinary parasitology 2015-09, Vol.212 (3-4), p.396-399
Hauptverfasser: Holt, Jennifer C., Purple, Kathryn E., Gerhold, Richard
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creator Holt, Jennifer C.
Purple, Kathryn E.
Gerhold, Richard
description •In this study, we evaluated non-indicating FTA Elute micro cards as a potential sampling substrate for the molecular identification of Trichomonas gallinae isolates.•Isolate concentrations of 10 or 100 trichomonads/40μl, from two T. gallinae isolates were inoculated onto a FTA Elute cards and analyzed by conventional polymerase chain reaction (PCR) using trichomonad-specific primers at 48h, 2 weeks, and 3 weeks post inoculation.•Three PCR-positive samples were detected at 48h from one isolate; however, all eluents from cards held for 2–3 weeks were PCR-negative. Our results suggest that FTA Elute cards have a low sensitivity for molecular identification of T. gallinae in low concentrations, such as those found in non-clinical birds; however, more research is needed to fully evaluate the efficacy of FTA Elute cards as a diagnostic tool for T. gallinae. Trichomonas gallinae is the causative agent for avian trichomonosis, which can have important population implications for domestic turkeys, columbids, raptors, and various passeriformes. Continued population surveillance and genotype distribution is needed to elucidate transmission dynamics and prevalence of T. gallinae among free-ranging birds. However, obtaining live cultures for laboratory testing is logistically challenging, limiting the ability to perform surveillance and genotype investigations. In this study, we evaluated non-indicating FTA Elute cards as a potential sampling storage substrate for downstream use in molecular identification of two T. gallinae isolates. Isolate concentrations of 10 or 100 trichomonads/40μl were inoculated onto a FTA Elute card in triplicate. At each time point (48h, 2 weeks, and 3 weeks), DNA elution procedures were performed on the cards, and the eluents were analyzed by conventional polymerase chain reaction (PCR) using trichomonad-specific primers. Three PCR-positive samples were detected at 48h from one isolate; however, all eluents from cards held for 2 and 3 weeks were PCR-negative. Our results suggest that use of FTA Elute cards for nucleic acid storage can lead to low PCR sensitivity of T. gallinae in low concentrations, such as those found in non-clinical birds; however, more research is needed to fully evaluate the efficacy of FTA Elute cards as a diagnostic tool for T. gallinae.
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Our results suggest that FTA Elute cards have a low sensitivity for molecular identification of T. gallinae in low concentrations, such as those found in non-clinical birds; however, more research is needed to fully evaluate the efficacy of FTA Elute cards as a diagnostic tool for T. gallinae. Trichomonas gallinae is the causative agent for avian trichomonosis, which can have important population implications for domestic turkeys, columbids, raptors, and various passeriformes. Continued population surveillance and genotype distribution is needed to elucidate transmission dynamics and prevalence of T. gallinae among free-ranging birds. However, obtaining live cultures for laboratory testing is logistically challenging, limiting the ability to perform surveillance and genotype investigations. In this study, we evaluated non-indicating FTA Elute cards as a potential sampling storage substrate for downstream use in molecular identification of two T. gallinae isolates. Isolate concentrations of 10 or 100 trichomonads/40μl were inoculated onto a FTA Elute card in triplicate. At each time point (48h, 2 weeks, and 3 weeks), DNA elution procedures were performed on the cards, and the eluents were analyzed by conventional polymerase chain reaction (PCR) using trichomonad-specific primers. Three PCR-positive samples were detected at 48h from one isolate; however, all eluents from cards held for 2 and 3 weeks were PCR-negative. Our results suggest that use of FTA Elute cards for nucleic acid storage can lead to low PCR sensitivity of T. gallinae in low concentrations, such as those found in non-clinical birds; however, more research is needed to fully evaluate the efficacy of FTA Elute cards as a diagnostic tool for T. gallinae.</description><identifier>ISSN: 0304-4017</identifier><identifier>EISSN: 1873-2550</identifier><identifier>DOI: 10.1016/j.vetpar.2015.07.017</identifier><identifier>PMID: 26228104</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Animals ; Avian ; Bird Diseases - parasitology ; DNA, Protozoan - genetics ; DNA, Protozoan - isolation &amp; purification ; Molecular sampling ; Nucleic Acids ; Specimen Handling - instrumentation ; Specimen Handling - methods ; Specimen Handling - veterinary ; Trichomonas - isolation &amp; purification ; Trichomonas gallinae ; Trichomonas Infections - diagnosis ; Trichomonas Infections - veterinary ; Trichomonosis</subject><ispartof>Veterinary parasitology, 2015-09, Vol.212 (3-4), p.396-399</ispartof><rights>2015 Elsevier B.V.</rights><rights>Copyright © 2015 Elsevier B.V. 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Our results suggest that FTA Elute cards have a low sensitivity for molecular identification of T. gallinae in low concentrations, such as those found in non-clinical birds; however, more research is needed to fully evaluate the efficacy of FTA Elute cards as a diagnostic tool for T. gallinae. Trichomonas gallinae is the causative agent for avian trichomonosis, which can have important population implications for domestic turkeys, columbids, raptors, and various passeriformes. Continued population surveillance and genotype distribution is needed to elucidate transmission dynamics and prevalence of T. gallinae among free-ranging birds. However, obtaining live cultures for laboratory testing is logistically challenging, limiting the ability to perform surveillance and genotype investigations. In this study, we evaluated non-indicating FTA Elute cards as a potential sampling storage substrate for downstream use in molecular identification of two T. gallinae isolates. Isolate concentrations of 10 or 100 trichomonads/40μl were inoculated onto a FTA Elute card in triplicate. At each time point (48h, 2 weeks, and 3 weeks), DNA elution procedures were performed on the cards, and the eluents were analyzed by conventional polymerase chain reaction (PCR) using trichomonad-specific primers. Three PCR-positive samples were detected at 48h from one isolate; however, all eluents from cards held for 2 and 3 weeks were PCR-negative. Our results suggest that use of FTA Elute cards for nucleic acid storage can lead to low PCR sensitivity of T. gallinae in low concentrations, such as those found in non-clinical birds; however, more research is needed to fully evaluate the efficacy of FTA Elute cards as a diagnostic tool for T. gallinae.</description><subject>Animals</subject><subject>Avian</subject><subject>Bird Diseases - parasitology</subject><subject>DNA, Protozoan - genetics</subject><subject>DNA, Protozoan - isolation &amp; purification</subject><subject>Molecular sampling</subject><subject>Nucleic Acids</subject><subject>Specimen Handling - instrumentation</subject><subject>Specimen Handling - methods</subject><subject>Specimen Handling - veterinary</subject><subject>Trichomonas - isolation &amp; purification</subject><subject>Trichomonas gallinae</subject><subject>Trichomonas Infections - diagnosis</subject><subject>Trichomonas Infections - veterinary</subject><subject>Trichomonosis</subject><issn>0304-4017</issn><issn>1873-2550</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE9LAzEQxYMotla_gcgevew6yf7JLoIgxapQ8NKeQzaZtCnbTU22hX57d2n16GngzXtvmB8h9xQSCrR42iQH7HbSJwxongBPgPILMqYlT2OW53BJxpBCFme9PiI3IWwAIIOCX5MRKxgrKWRj8rwMGDkTzRavUYdq3brGrY6RcT7S2Audde2wX3ir1m7rWhmilWwa20q8JVdGNgHvznNClrO3xfQjnn-9f05f57FKC9bFuQKmTVnWJudoADhPFRomlYaq4oWkdYkoU81rXVdlnZtSUdBa1rpilcqzdEIeT7077773GDqxtUFh08gW3T4IyulQVJVVb81OVuVdCB6N2Hm7lf4oKIgBm9iIEzYxYBPARY-njz2cL-zrLeq_0C-n3vByMmD_58GiF0FZbBVq63tIQjv7_4UfLtqAcQ</recordid><startdate>20150915</startdate><enddate>20150915</enddate><creator>Holt, Jennifer C.</creator><creator>Purple, Kathryn E.</creator><creator>Gerhold, Richard</creator><general>Elsevier B.V</general><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>20150915</creationdate><title>Use of FTA technology for detection of Trichomonas gallinae</title><author>Holt, Jennifer C. ; Purple, Kathryn E. ; Gerhold, Richard</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c362t-5c02df88bf57ef00773cef2acd09976a1b8eea3d7bdb98b5f8c10ddabd929c543</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Animals</topic><topic>Avian</topic><topic>Bird Diseases - parasitology</topic><topic>DNA, Protozoan - genetics</topic><topic>DNA, Protozoan - isolation &amp; purification</topic><topic>Molecular sampling</topic><topic>Nucleic Acids</topic><topic>Specimen Handling - instrumentation</topic><topic>Specimen Handling - methods</topic><topic>Specimen Handling - veterinary</topic><topic>Trichomonas - isolation &amp; purification</topic><topic>Trichomonas gallinae</topic><topic>Trichomonas Infections - diagnosis</topic><topic>Trichomonas Infections - veterinary</topic><topic>Trichomonosis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Holt, Jennifer C.</creatorcontrib><creatorcontrib>Purple, Kathryn E.</creatorcontrib><creatorcontrib>Gerhold, Richard</creatorcontrib><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>Veterinary parasitology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Holt, Jennifer C.</au><au>Purple, Kathryn E.</au><au>Gerhold, Richard</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Use of FTA technology for detection of Trichomonas gallinae</atitle><jtitle>Veterinary parasitology</jtitle><addtitle>Vet Parasitol</addtitle><date>2015-09-15</date><risdate>2015</risdate><volume>212</volume><issue>3-4</issue><spage>396</spage><epage>399</epage><pages>396-399</pages><issn>0304-4017</issn><eissn>1873-2550</eissn><abstract>•In this study, we evaluated non-indicating FTA Elute micro cards as a potential sampling substrate for the molecular identification of Trichomonas gallinae isolates.•Isolate concentrations of 10 or 100 trichomonads/40μl, from two T. gallinae isolates were inoculated onto a FTA Elute cards and analyzed by conventional polymerase chain reaction (PCR) using trichomonad-specific primers at 48h, 2 weeks, and 3 weeks post inoculation.•Three PCR-positive samples were detected at 48h from one isolate; however, all eluents from cards held for 2–3 weeks were PCR-negative. 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Isolate concentrations of 10 or 100 trichomonads/40μl were inoculated onto a FTA Elute card in triplicate. At each time point (48h, 2 weeks, and 3 weeks), DNA elution procedures were performed on the cards, and the eluents were analyzed by conventional polymerase chain reaction (PCR) using trichomonad-specific primers. Three PCR-positive samples were detected at 48h from one isolate; however, all eluents from cards held for 2 and 3 weeks were PCR-negative. Our results suggest that use of FTA Elute cards for nucleic acid storage can lead to low PCR sensitivity of T. gallinae in low concentrations, such as those found in non-clinical birds; however, more research is needed to fully evaluate the efficacy of FTA Elute cards as a diagnostic tool for T. gallinae.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>26228104</pmid><doi>10.1016/j.vetpar.2015.07.017</doi><tpages>4</tpages></addata></record>
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subjects Animals
Avian
Bird Diseases - parasitology
DNA, Protozoan - genetics
DNA, Protozoan - isolation & purification
Molecular sampling
Nucleic Acids
Specimen Handling - instrumentation
Specimen Handling - methods
Specimen Handling - veterinary
Trichomonas - isolation & purification
Trichomonas gallinae
Trichomonas Infections - diagnosis
Trichomonas Infections - veterinary
Trichomonosis
title Use of FTA technology for detection of Trichomonas gallinae
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