Development of specific and rapid detection of bacterial pathogens in dairy products by PCR
A simple and specific method for direct detection of bovine mastitis pathogens (Streptococcus agalactiae (GBS), Staphylococcus aureus and Escherichia coli) in milk products, bacterial samples from milk and isolated bacterial DNA was developed. The method is based on PCR using sequence-specific prime...
Gespeichert in:
| Veröffentlicht in: | Folia microbiologica 2006-01, Vol.51 (6), p.639-646 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 646 |
|---|---|
| container_issue | 6 |
| container_start_page | 639 |
| container_title | Folia microbiologica |
| container_volume | 51 |
| creator | Chotar, M Vidova, B Godany, A |
| description | A simple and specific method for direct detection of bovine mastitis pathogens (Streptococcus agalactiae (GBS), Staphylococcus aureus and Escherichia coli) in milk products, bacterial samples from milk and isolated bacterial DNA was developed. The method is based on PCR using sequence-specific primers only for GBS and species-specific primers derived from 16S and 23S rRNA for all chosen species. The presence of the gene of surface immunogenic protein (Sip) in bovine GBS isolates was confirmed. The GBS detection was performed with the sequence coding for surface immunogenic protein from GBS human isolates designated as Sip specific sequence (SSS); this sequence was selected for specific primer design. The sequence is unique for GBS and was designed from a consensus of all known sip genes. The specific identification was shown on a collection of 75 GBS bovine isolates from different localities in Slovakia. All isolates were positive to SSS, 16S and 23S rRNA sequence. The 16S and 23S rRNA PCR detection was also performed with S. aureus and E. coli isolates and specific PCR products were also detected. The detection limit of this assay for milk products was 6 CFU/microL for GBS and E. coli, and 16 CFU/microL for S. aureus. This rapid, sensitive and specific diagnostic method can be performed within hours and represents an innovative diagnostic tool for the detection of milk pathogens in dairy products. |
| doi_str_mv | 10.1007/BF02931632 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_19589773</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>19589773</sourcerecordid><originalsourceid>FETCH-LOGICAL-c398t-49031df66e372686c4958e6591aeff3e703f00d5ecc71f9c7babb0137f547e73</originalsourceid><addsrcrecordid>eNp9kU2LFDEQhoMo7uzqxbsSPIgIrZXOV-eos64KC4rsSQ8hna6sWXo6bdItzL83wwwsePBUUPXwFFUvIc8YvGUA-t2HK2gNZ4q3D8iGdVo0hkv1kGwAmGxk7Z-R81LuABQI3j4mZ0wLKTsQG_LzEv_gmOYdTgtNgZYZfQzRUzcNNLs5DnTABf0S03SY984vmKMb6eyWX-kWp0LjRAcX857OOQ2rXwrt9_Tb9vsT8ii4seDTU70gN1cfb7afm-uvn75s3183nptuaYQBzoagFHLdqk55YWSHShrmMASOGngAGCR6r1kwXveu74FxHaTQqPkFeXXU1vW_VyyL3cXicRzdhGktllWd0ZpX8PX_Qc6kYtByVdGX_6B3ac1TvcJqKYyovzv43hwhn1MpGYOdc9y5vLcM7CEZe59MhV-cjGu_w-EePUVRgedHILhk3W2OxW5_tNUCNUbT8b8l0Y9_</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>754948043</pqid></control><display><type>article</type><title>Development of specific and rapid detection of bacterial pathogens in dairy products by PCR</title><source>MEDLINE</source><source>SpringerNature Journals</source><creator>Chotar, M ; Vidova, B ; Godany, A</creator><creatorcontrib>Chotar, M ; Vidova, B ; Godany, A</creatorcontrib><description>A simple and specific method for direct detection of bovine mastitis pathogens (Streptococcus agalactiae (GBS), Staphylococcus aureus and Escherichia coli) in milk products, bacterial samples from milk and isolated bacterial DNA was developed. The method is based on PCR using sequence-specific primers only for GBS and species-specific primers derived from 16S and 23S rRNA for all chosen species. The presence of the gene of surface immunogenic protein (Sip) in bovine GBS isolates was confirmed. The GBS detection was performed with the sequence coding for surface immunogenic protein from GBS human isolates designated as Sip specific sequence (SSS); this sequence was selected for specific primer design. The sequence is unique for GBS and was designed from a consensus of all known sip genes. The specific identification was shown on a collection of 75 GBS bovine isolates from different localities in Slovakia. All isolates were positive to SSS, 16S and 23S rRNA sequence. The 16S and 23S rRNA PCR detection was also performed with S. aureus and E. coli isolates and specific PCR products were also detected. The detection limit of this assay for milk products was 6 CFU/microL for GBS and E. coli, and 16 CFU/microL for S. aureus. This rapid, sensitive and specific diagnostic method can be performed within hours and represents an innovative diagnostic tool for the detection of milk pathogens in dairy products.</description><identifier>ISSN: 0015-5632</identifier><identifier>EISSN: 1874-9356</identifier><identifier>DOI: 10.1007/BF02931632</identifier><identifier>PMID: 17455804</identifier><language>eng</language><publisher>United States: Springer Nature B.V</publisher><subject>ADN ; AGENT PATHOGENE ; AISLAMIENTO ; Antigens, Bacterial - genetics ; Bacteria ; Bacteriological Techniques ; Base Sequence ; Colony-forming cells ; Conserved sequence ; COW MILK ; Dairy products ; Dairy Products - microbiology ; DNA ; DNA Primers ; DNA, Bacterial - analysis ; DNA, Bacterial - genetics ; DNA, Ribosomal - analysis ; DNA, Ribosomal - genetics ; Electrophoresis, Agar Gel ; ESCHERICHIA COLI ; Escherichia coli - genetics ; Escherichia coli - isolation & purification ; Food Microbiology ; GENE ; GENES ; Geographical variations ; http://www.fao.org/aos/agrovoc#c_16080 ; http://www.fao.org/aos/agrovoc#c_16429 ; http://www.fao.org/aos/agrovoc#c_2347 ; http://www.fao.org/aos/agrovoc#c_26700 ; http://www.fao.org/aos/agrovoc#c_27583 ; http://www.fao.org/aos/agrovoc#c_3214 ; http://www.fao.org/aos/agrovoc#c_33700 ; http://www.fao.org/aos/agrovoc#c_34079 ; http://www.fao.org/aos/agrovoc#c_37744 ; http://www.fao.org/aos/agrovoc#c_4788 ; http://www.fao.org/aos/agrovoc#c_4830 ; http://www.fao.org/aos/agrovoc#c_5630 ; Immunogenicity ; ISOLATION ; ISOLEMENT ; LAIT DE VACHE ; LECHE DE VACA ; Mastitis ; METHODE ; METHODS ; METODOS ; Microbiology ; Milk ; MILK PRODUCTS ; Molecular Sequence Data ; NUCLEOTIDE SEQUENCE ; ORGANISMOS PATOGENOS ; PATHOGENS ; PCR ; Polymerase chain reaction ; Polymerase Chain Reaction - methods ; Primers ; PRODUCTOS LACTEOS ; PRODUIT LAITIER ; RNA, Ribosomal, 16S - genetics ; RNA, Ribosomal, 23S - genetics ; rRNA 16S ; rRNA 23S ; SECUENCIA NUCLEOTIDICA ; Sensitivity and Specificity ; SEQUENCE NUCLEOTIDIQUE ; STAPHYLOCOCCUS AUREAS ; STAPHYLOCOCCUS AUREUS ; Staphylococcus aureus - genetics ; Staphylococcus aureus - isolation & purification ; STREPTOCOCCUS AGALACTIAE ; Streptococcus agalactiae - genetics ; Streptococcus agalactiae - isolation & purification</subject><ispartof>Folia microbiologica, 2006-01, Vol.51 (6), p.639-646</ispartof><rights>Institute of Microbiology, Academy of Sciences of the Czech Republic 2006</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c398t-49031df66e372686c4958e6591aeff3e703f00d5ecc71f9c7babb0137f547e73</citedby><cites>FETCH-LOGICAL-c398t-49031df66e372686c4958e6591aeff3e703f00d5ecc71f9c7babb0137f547e73</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17455804$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chotar, M</creatorcontrib><creatorcontrib>Vidova, B</creatorcontrib><creatorcontrib>Godany, A</creatorcontrib><title>Development of specific and rapid detection of bacterial pathogens in dairy products by PCR</title><title>Folia microbiologica</title><addtitle>Folia Microbiol (Praha)</addtitle><description>A simple and specific method for direct detection of bovine mastitis pathogens (Streptococcus agalactiae (GBS), Staphylococcus aureus and Escherichia coli) in milk products, bacterial samples from milk and isolated bacterial DNA was developed. The method is based on PCR using sequence-specific primers only for GBS and species-specific primers derived from 16S and 23S rRNA for all chosen species. The presence of the gene of surface immunogenic protein (Sip) in bovine GBS isolates was confirmed. The GBS detection was performed with the sequence coding for surface immunogenic protein from GBS human isolates designated as Sip specific sequence (SSS); this sequence was selected for specific primer design. The sequence is unique for GBS and was designed from a consensus of all known sip genes. The specific identification was shown on a collection of 75 GBS bovine isolates from different localities in Slovakia. All isolates were positive to SSS, 16S and 23S rRNA sequence. The 16S and 23S rRNA PCR detection was also performed with S. aureus and E. coli isolates and specific PCR products were also detected. The detection limit of this assay for milk products was 6 CFU/microL for GBS and E. coli, and 16 CFU/microL for S. aureus. This rapid, sensitive and specific diagnostic method can be performed within hours and represents an innovative diagnostic tool for the detection of milk pathogens in dairy products.</description><subject>ADN</subject><subject>AGENT PATHOGENE</subject><subject>AISLAMIENTO</subject><subject>Antigens, Bacterial - genetics</subject><subject>Bacteria</subject><subject>Bacteriological Techniques</subject><subject>Base Sequence</subject><subject>Colony-forming cells</subject><subject>Conserved sequence</subject><subject>COW MILK</subject><subject>Dairy products</subject><subject>Dairy Products - microbiology</subject><subject>DNA</subject><subject>DNA Primers</subject><subject>DNA, Bacterial - analysis</subject><subject>DNA, Bacterial - genetics</subject><subject>DNA, Ribosomal - analysis</subject><subject>DNA, Ribosomal - genetics</subject><subject>Electrophoresis, Agar Gel</subject><subject>ESCHERICHIA COLI</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - isolation & purification</subject><subject>Food Microbiology</subject><subject>GENE</subject><subject>GENES</subject><subject>Geographical variations</subject><subject>http://www.fao.org/aos/agrovoc#c_16080</subject><subject>http://www.fao.org/aos/agrovoc#c_16429</subject><subject>http://www.fao.org/aos/agrovoc#c_2347</subject><subject>http://www.fao.org/aos/agrovoc#c_26700</subject><subject>http://www.fao.org/aos/agrovoc#c_27583</subject><subject>http://www.fao.org/aos/agrovoc#c_3214</subject><subject>http://www.fao.org/aos/agrovoc#c_33700</subject><subject>http://www.fao.org/aos/agrovoc#c_34079</subject><subject>http://www.fao.org/aos/agrovoc#c_37744</subject><subject>http://www.fao.org/aos/agrovoc#c_4788</subject><subject>http://www.fao.org/aos/agrovoc#c_4830</subject><subject>http://www.fao.org/aos/agrovoc#c_5630</subject><subject>Immunogenicity</subject><subject>ISOLATION</subject><subject>ISOLEMENT</subject><subject>LAIT DE VACHE</subject><subject>LECHE DE VACA</subject><subject>Mastitis</subject><subject>METHODE</subject><subject>METHODS</subject><subject>METODOS</subject><subject>Microbiology</subject><subject>Milk</subject><subject>MILK PRODUCTS</subject><subject>Molecular Sequence Data</subject><subject>NUCLEOTIDE SEQUENCE</subject><subject>ORGANISMOS PATOGENOS</subject><subject>PATHOGENS</subject><subject>PCR</subject><subject>Polymerase chain reaction</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Primers</subject><subject>PRODUCTOS LACTEOS</subject><subject>PRODUIT LAITIER</subject><subject>RNA, Ribosomal, 16S - genetics</subject><subject>RNA, Ribosomal, 23S - genetics</subject><subject>rRNA 16S</subject><subject>rRNA 23S</subject><subject>SECUENCIA NUCLEOTIDICA</subject><subject>Sensitivity and Specificity</subject><subject>SEQUENCE NUCLEOTIDIQUE</subject><subject>STAPHYLOCOCCUS AUREAS</subject><subject>STAPHYLOCOCCUS AUREUS</subject><subject>Staphylococcus aureus - genetics</subject><subject>Staphylococcus aureus - isolation & purification</subject><subject>STREPTOCOCCUS AGALACTIAE</subject><subject>Streptococcus agalactiae - genetics</subject><subject>Streptococcus agalactiae - isolation & purification</subject><issn>0015-5632</issn><issn>1874-9356</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kU2LFDEQhoMo7uzqxbsSPIgIrZXOV-eos64KC4rsSQ8hna6sWXo6bdItzL83wwwsePBUUPXwFFUvIc8YvGUA-t2HK2gNZ4q3D8iGdVo0hkv1kGwAmGxk7Z-R81LuABQI3j4mZ0wLKTsQG_LzEv_gmOYdTgtNgZYZfQzRUzcNNLs5DnTABf0S03SY984vmKMb6eyWX-kWp0LjRAcX857OOQ2rXwrt9_Tb9vsT8ii4seDTU70gN1cfb7afm-uvn75s3183nptuaYQBzoagFHLdqk55YWSHShrmMASOGngAGCR6r1kwXveu74FxHaTQqPkFeXXU1vW_VyyL3cXicRzdhGktllWd0ZpX8PX_Qc6kYtByVdGX_6B3ac1TvcJqKYyovzv43hwhn1MpGYOdc9y5vLcM7CEZe59MhV-cjGu_w-EePUVRgedHILhk3W2OxW5_tNUCNUbT8b8l0Y9_</recordid><startdate>20060101</startdate><enddate>20060101</enddate><creator>Chotar, M</creator><creator>Vidova, B</creator><creator>Godany, A</creator><general>Springer Nature B.V</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>3V.</scope><scope>7QL</scope><scope>7T7</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope></search><sort><creationdate>20060101</creationdate><title>Development of specific and rapid detection of bacterial pathogens in dairy products by PCR</title><author>Chotar, M ; Vidova, B ; Godany, A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c398t-49031df66e372686c4958e6591aeff3e703f00d5ecc71f9c7babb0137f547e73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>ADN</topic><topic>AGENT PATHOGENE</topic><topic>AISLAMIENTO</topic><topic>Antigens, Bacterial - genetics</topic><topic>Bacteria</topic><topic>Bacteriological Techniques</topic><topic>Base Sequence</topic><topic>Colony-forming cells</topic><topic>Conserved sequence</topic><topic>COW MILK</topic><topic>Dairy products</topic><topic>Dairy Products - microbiology</topic><topic>DNA</topic><topic>DNA Primers</topic><topic>DNA, Bacterial - analysis</topic><topic>DNA, Bacterial - genetics</topic><topic>DNA, Ribosomal - analysis</topic><topic>DNA, Ribosomal - genetics</topic><topic>Electrophoresis, Agar Gel</topic><topic>ESCHERICHIA COLI</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - isolation & purification</topic><topic>Food Microbiology</topic><topic>GENE</topic><topic>GENES</topic><topic>Geographical variations</topic><topic>http://www.fao.org/aos/agrovoc#c_16080</topic><topic>http://www.fao.org/aos/agrovoc#c_16429</topic><topic>http://www.fao.org/aos/agrovoc#c_2347</topic><topic>http://www.fao.org/aos/agrovoc#c_26700</topic><topic>http://www.fao.org/aos/agrovoc#c_27583</topic><topic>http://www.fao.org/aos/agrovoc#c_3214</topic><topic>http://www.fao.org/aos/agrovoc#c_33700</topic><topic>http://www.fao.org/aos/agrovoc#c_34079</topic><topic>http://www.fao.org/aos/agrovoc#c_37744</topic><topic>http://www.fao.org/aos/agrovoc#c_4788</topic><topic>http://www.fao.org/aos/agrovoc#c_4830</topic><topic>http://www.fao.org/aos/agrovoc#c_5630</topic><topic>Immunogenicity</topic><topic>ISOLATION</topic><topic>ISOLEMENT</topic><topic>LAIT DE VACHE</topic><topic>LECHE DE VACA</topic><topic>Mastitis</topic><topic>METHODE</topic><topic>METHODS</topic><topic>METODOS</topic><topic>Microbiology</topic><topic>Milk</topic><topic>MILK PRODUCTS</topic><topic>Molecular Sequence Data</topic><topic>NUCLEOTIDE SEQUENCE</topic><topic>ORGANISMOS PATOGENOS</topic><topic>PATHOGENS</topic><topic>PCR</topic><topic>Polymerase chain reaction</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Primers</topic><topic>PRODUCTOS LACTEOS</topic><topic>PRODUIT LAITIER</topic><topic>RNA, Ribosomal, 16S - genetics</topic><topic>RNA, Ribosomal, 23S - genetics</topic><topic>rRNA 16S</topic><topic>rRNA 23S</topic><topic>SECUENCIA NUCLEOTIDICA</topic><topic>Sensitivity and Specificity</topic><topic>SEQUENCE NUCLEOTIDIQUE</topic><topic>STAPHYLOCOCCUS AUREAS</topic><topic>STAPHYLOCOCCUS AUREUS</topic><topic>Staphylococcus aureus - genetics</topic><topic>Staphylococcus aureus - isolation & purification</topic><topic>STREPTOCOCCUS AGALACTIAE</topic><topic>Streptococcus agalactiae - genetics</topic><topic>Streptococcus agalactiae - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chotar, M</creatorcontrib><creatorcontrib>Vidova, B</creatorcontrib><creatorcontrib>Godany, A</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>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><jtitle>Folia microbiologica</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chotar, M</au><au>Vidova, B</au><au>Godany, A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of specific and rapid detection of bacterial pathogens in dairy products by PCR</atitle><jtitle>Folia microbiologica</jtitle><addtitle>Folia Microbiol (Praha)</addtitle><date>2006-01-01</date><risdate>2006</risdate><volume>51</volume><issue>6</issue><spage>639</spage><epage>646</epage><pages>639-646</pages><issn>0015-5632</issn><eissn>1874-9356</eissn><abstract>A simple and specific method for direct detection of bovine mastitis pathogens (Streptococcus agalactiae (GBS), Staphylococcus aureus and Escherichia coli) in milk products, bacterial samples from milk and isolated bacterial DNA was developed. The method is based on PCR using sequence-specific primers only for GBS and species-specific primers derived from 16S and 23S rRNA for all chosen species. The presence of the gene of surface immunogenic protein (Sip) in bovine GBS isolates was confirmed. The GBS detection was performed with the sequence coding for surface immunogenic protein from GBS human isolates designated as Sip specific sequence (SSS); this sequence was selected for specific primer design. The sequence is unique for GBS and was designed from a consensus of all known sip genes. The specific identification was shown on a collection of 75 GBS bovine isolates from different localities in Slovakia. All isolates were positive to SSS, 16S and 23S rRNA sequence. The 16S and 23S rRNA PCR detection was also performed with S. aureus and E. coli isolates and specific PCR products were also detected. The detection limit of this assay for milk products was 6 CFU/microL for GBS and E. coli, and 16 CFU/microL for S. aureus. This rapid, sensitive and specific diagnostic method can be performed within hours and represents an innovative diagnostic tool for the detection of milk pathogens in dairy products.</abstract><cop>United States</cop><pub>Springer Nature B.V</pub><pmid>17455804</pmid><doi>10.1007/BF02931632</doi><tpages>8</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0015-5632 |
| ispartof | Folia microbiologica, 2006-01, Vol.51 (6), p.639-646 |
| issn | 0015-5632 1874-9356 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_19589773 |
| source | MEDLINE; SpringerNature Journals |
| subjects | ADN AGENT PATHOGENE AISLAMIENTO Antigens, Bacterial - genetics Bacteria Bacteriological Techniques Base Sequence Colony-forming cells Conserved sequence COW MILK Dairy products Dairy Products - microbiology DNA DNA Primers DNA, Bacterial - analysis DNA, Bacterial - genetics DNA, Ribosomal - analysis DNA, Ribosomal - genetics Electrophoresis, Agar Gel ESCHERICHIA COLI Escherichia coli - genetics Escherichia coli - isolation & purification Food Microbiology GENE GENES Geographical variations http://www.fao.org/aos/agrovoc#c_16080 http://www.fao.org/aos/agrovoc#c_16429 http://www.fao.org/aos/agrovoc#c_2347 http://www.fao.org/aos/agrovoc#c_26700 http://www.fao.org/aos/agrovoc#c_27583 http://www.fao.org/aos/agrovoc#c_3214 http://www.fao.org/aos/agrovoc#c_33700 http://www.fao.org/aos/agrovoc#c_34079 http://www.fao.org/aos/agrovoc#c_37744 http://www.fao.org/aos/agrovoc#c_4788 http://www.fao.org/aos/agrovoc#c_4830 http://www.fao.org/aos/agrovoc#c_5630 Immunogenicity ISOLATION ISOLEMENT LAIT DE VACHE LECHE DE VACA Mastitis METHODE METHODS METODOS Microbiology Milk MILK PRODUCTS Molecular Sequence Data NUCLEOTIDE SEQUENCE ORGANISMOS PATOGENOS PATHOGENS PCR Polymerase chain reaction Polymerase Chain Reaction - methods Primers PRODUCTOS LACTEOS PRODUIT LAITIER RNA, Ribosomal, 16S - genetics RNA, Ribosomal, 23S - genetics rRNA 16S rRNA 23S SECUENCIA NUCLEOTIDICA Sensitivity and Specificity SEQUENCE NUCLEOTIDIQUE STAPHYLOCOCCUS AUREAS STAPHYLOCOCCUS AUREUS Staphylococcus aureus - genetics Staphylococcus aureus - isolation & purification STREPTOCOCCUS AGALACTIAE Streptococcus agalactiae - genetics Streptococcus agalactiae - isolation & purification |
| title | Development of specific and rapid detection of bacterial pathogens in dairy products by PCR |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-03T22%3A28%3A02IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Development%20of%20specific%20and%20rapid%20detection%20of%20bacterial%20pathogens%20in%20dairy%20products%20by%20PCR&rft.jtitle=Folia%20microbiologica&rft.au=Chotar,%20M&rft.date=2006-01-01&rft.volume=51&rft.issue=6&rft.spage=639&rft.epage=646&rft.pages=639-646&rft.issn=0015-5632&rft.eissn=1874-9356&rft_id=info:doi/10.1007/BF02931632&rft_dat=%3Cproquest_cross%3E19589773%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=754948043&rft_id=info:pmid/17455804&rfr_iscdi=true |