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Quantification of infectious Human mastadenovirus in environmental matrices using PMAxx-qPCR
Molecular methodologies providing data on viral concentration and infectivity have been successfully used in environmental virology, supporting quantitative risk assessment studies. The present study aimed to assess human mastadenovirus (HAdV) intact particles using a derivative of propidium monoazi...
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| Published in: | Brazilian journal of microbiology 2022-09, Vol.53 (3), p.1465-1471 |
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| Main Authors: | , , , , , , |
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| cites | cdi_FETCH-LOGICAL-c474t-5cfef82ddc3321b49d30601bc7ea0f3c7056b3abc4b365d431ca8976c785765c3 |
| container_end_page | 1471 |
| container_issue | 3 |
| container_start_page | 1465 |
| container_title | Brazilian journal of microbiology |
| container_volume | 53 |
| creator | Pedrosa de Macena, Lorena da Graça Pereira, Joseane Simone de Oliveira da Silva, Jansen Couto Ferreira, Fernando César Maranhão, Adriana Gonçalves Lanzarini, Natália Maria Miagostovich, Marize Pereira |
| description | Molecular methodologies providing data on viral concentration and infectivity have been successfully used in environmental virology, supporting quantitative risk assessment studies. The present study aimed to assess
human mastadenovirus
(HAdV) intact particles using a derivative of propidium monoazide associated with qPCR (PMAxx-qPCR) in aquatic matrices. Initially, different concentrations of PMAxx were evaluated to establish an optimal protocol for treating different naturally contaminated matrices, using 10 min incubation in the dark at 200 rpm at room temperature and 15 min of photoactivation in the PMA-Lite™ LED photolysis device. There was no significant reduction in the quantification of infectious HAdV with increasing concentration of PMAxx used (20 μM, 50 μM, and 100 μM), except for sewage samples. In this matrix, a reduction of 5.01 log of genomic copies (GC)/L was observed from the concentration of 50 μM and revealed 100% HAdV particles with damaged capsids. On the other hand, the mean reduction of 0.51 log in stool samples using the same concentration mentioned above demonstrated 83% of damaged particles eliminated in the stool. Following, 50 μM PMAxx-qPCR protocol revealed a log reduction of 0.91, 0.67, and 1.05 in other samples of raw sewage, brackish, and seawater where HAdV concentration reached 1.47 × 10
4
, 6.81 × 10
2
, and 2.33 × 10
2
GC/L, respectively. Fifty micrometers of PMAxx protocol helped screen intact viruses from different matrices, including sea and brackish water. |
| doi_str_mv | 10.1007/s42770-022-00775-5 |
| format | article |
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human mastadenovirus
(HAdV) intact particles using a derivative of propidium monoazide associated with qPCR (PMAxx-qPCR) in aquatic matrices. Initially, different concentrations of PMAxx were evaluated to establish an optimal protocol for treating different naturally contaminated matrices, using 10 min incubation in the dark at 200 rpm at room temperature and 15 min of photoactivation in the PMA-Lite™ LED photolysis device. There was no significant reduction in the quantification of infectious HAdV with increasing concentration of PMAxx used (20 μM, 50 μM, and 100 μM), except for sewage samples. In this matrix, a reduction of 5.01 log of genomic copies (GC)/L was observed from the concentration of 50 μM and revealed 100% HAdV particles with damaged capsids. On the other hand, the mean reduction of 0.51 log in stool samples using the same concentration mentioned above demonstrated 83% of damaged particles eliminated in the stool. Following, 50 μM PMAxx-qPCR protocol revealed a log reduction of 0.91, 0.67, and 1.05 in other samples of raw sewage, brackish, and seawater where HAdV concentration reached 1.47 × 10
4
, 6.81 × 10
2
, and 2.33 × 10
2
GC/L, respectively. Fifty micrometers of PMAxx protocol helped screen intact viruses from different matrices, including sea and brackish water.</description><identifier>ISSN: 1517-8382</identifier><identifier>EISSN: 1678-4405</identifier><identifier>DOI: 10.1007/s42770-022-00775-5</identifier><identifier>PMID: 35666431</identifier><language>eng</language><publisher>Cham: Springer International Publishing</publisher><subject>Biomedical and Life Sciences ; Brackish water ; Capsids ; Chemical analysis ; Environmental Microbiology - Research Paper ; Food Microbiology ; Infectivity ; Life Sciences ; Medical Microbiology ; Microbial Ecology ; Microbial Genetics and Genomics ; Microbiology ; Micrometers ; Mycology ; Photoactivation ; Photolysis ; Raw sewage ; Reduction ; Risk assessment ; Room temperature ; Seawater ; Sewage ; Viruses ; Water analysis</subject><ispartof>Brazilian journal of microbiology, 2022-09, Vol.53 (3), p.1465-1471</ispartof><rights>The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia 2022</rights><rights>2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.</rights><rights>The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia 2022.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c474t-5cfef82ddc3321b49d30601bc7ea0f3c7056b3abc4b365d431ca8976c785765c3</citedby><cites>FETCH-LOGICAL-c474t-5cfef82ddc3321b49d30601bc7ea0f3c7056b3abc4b365d431ca8976c785765c3</cites><orcidid>0000-0003-1420-6010</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9168632/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9168632/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35666431$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pedrosa de Macena, Lorena da Graça</creatorcontrib><creatorcontrib>Pereira, Joseane Simone de Oliveira</creatorcontrib><creatorcontrib>da Silva, Jansen Couto</creatorcontrib><creatorcontrib>Ferreira, Fernando César</creatorcontrib><creatorcontrib>Maranhão, Adriana Gonçalves</creatorcontrib><creatorcontrib>Lanzarini, Natália Maria</creatorcontrib><creatorcontrib>Miagostovich, Marize Pereira</creatorcontrib><title>Quantification of infectious Human mastadenovirus in environmental matrices using PMAxx-qPCR</title><title>Brazilian journal of microbiology</title><addtitle>Braz J Microbiol</addtitle><addtitle>Braz J Microbiol</addtitle><description>Molecular methodologies providing data on viral concentration and infectivity have been successfully used in environmental virology, supporting quantitative risk assessment studies. The present study aimed to assess
human mastadenovirus
(HAdV) intact particles using a derivative of propidium monoazide associated with qPCR (PMAxx-qPCR) in aquatic matrices. Initially, different concentrations of PMAxx were evaluated to establish an optimal protocol for treating different naturally contaminated matrices, using 10 min incubation in the dark at 200 rpm at room temperature and 15 min of photoactivation in the PMA-Lite™ LED photolysis device. There was no significant reduction in the quantification of infectious HAdV with increasing concentration of PMAxx used (20 μM, 50 μM, and 100 μM), except for sewage samples. In this matrix, a reduction of 5.01 log of genomic copies (GC)/L was observed from the concentration of 50 μM and revealed 100% HAdV particles with damaged capsids. On the other hand, the mean reduction of 0.51 log in stool samples using the same concentration mentioned above demonstrated 83% of damaged particles eliminated in the stool. Following, 50 μM PMAxx-qPCR protocol revealed a log reduction of 0.91, 0.67, and 1.05 in other samples of raw sewage, brackish, and seawater where HAdV concentration reached 1.47 × 10
4
, 6.81 × 10
2
, and 2.33 × 10
2
GC/L, respectively. Fifty micrometers of PMAxx protocol helped screen intact viruses from different matrices, including sea and brackish water.</description><subject>Biomedical and Life Sciences</subject><subject>Brackish water</subject><subject>Capsids</subject><subject>Chemical analysis</subject><subject>Environmental Microbiology - Research Paper</subject><subject>Food Microbiology</subject><subject>Infectivity</subject><subject>Life Sciences</subject><subject>Medical Microbiology</subject><subject>Microbial Ecology</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Micrometers</subject><subject>Mycology</subject><subject>Photoactivation</subject><subject>Photolysis</subject><subject>Raw sewage</subject><subject>Reduction</subject><subject>Risk assessment</subject><subject>Room temperature</subject><subject>Seawater</subject><subject>Sewage</subject><subject>Viruses</subject><subject>Water analysis</subject><issn>1517-8382</issn><issn>1678-4405</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><recordid>eNp9UV1rFTEQDUVpa-0f6IMs-OJLdPK990UoF7VCxVb0rRCy2ew1ZTdpk91S_72jt9bqQyGQmcyZM2dyCDli8JoBmDdVcmOAAucUU6Oo2iH7TJuWSgnqCcaKGdqKlu-RZ7VeAnAFku-SPaG01lKwfXJxvrg0xyF6N8ecmjw0MQ3BY7LU5mSZXGomV2fXh5RvYsHHmJqQMMxpCml2I9bnEn2ozVJj2jRnn45vb-n12frLc_J0cGMNh3f3Afn2_t3X9Qk9_fzh4_r4lHpp5EyVH8LQ8r73QnDWyVUvQAPrvAkOBuENKN0J13nZCa161O1duzLam1YZrbw4IG-3vFdLN4Xeo6ziRntV4uTKD5tdtP9WUvxuN_nGrphuteBI8OqOoOTrJdTZTrH6MI4uBfwHy7WRAHhWCH35H_QyLyXhepYbaI2WWgOi-BblS661hOFeDAP7yzy7Nc-iefa3eVZh04uHa9y3_HELAWILqFhKm1D-zn6E9ifB76bB</recordid><startdate>20220901</startdate><enddate>20220901</enddate><creator>Pedrosa de Macena, Lorena da Graça</creator><creator>Pereira, Joseane Simone de Oliveira</creator><creator>da Silva, Jansen Couto</creator><creator>Ferreira, Fernando César</creator><creator>Maranhão, Adriana Gonçalves</creator><creator>Lanzarini, Natália Maria</creator><creator>Miagostovich, Marize Pereira</creator><general>Springer International Publishing</general><general>Springer Nature B.V</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7U9</scope><scope>C1K</scope><scope>H94</scope><scope>K9.</scope><scope>M7N</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0003-1420-6010</orcidid></search><sort><creationdate>20220901</creationdate><title>Quantification of infectious Human mastadenovirus in environmental matrices using PMAxx-qPCR</title><author>Pedrosa de Macena, Lorena da Graça ; Pereira, Joseane Simone de Oliveira ; da Silva, Jansen Couto ; Ferreira, Fernando César ; Maranhão, Adriana Gonçalves ; Lanzarini, Natália Maria ; Miagostovich, Marize Pereira</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c474t-5cfef82ddc3321b49d30601bc7ea0f3c7056b3abc4b365d431ca8976c785765c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Biomedical and Life Sciences</topic><topic>Brackish water</topic><topic>Capsids</topic><topic>Chemical analysis</topic><topic>Environmental Microbiology - Research Paper</topic><topic>Food Microbiology</topic><topic>Infectivity</topic><topic>Life Sciences</topic><topic>Medical Microbiology</topic><topic>Microbial Ecology</topic><topic>Microbial Genetics and Genomics</topic><topic>Microbiology</topic><topic>Micrometers</topic><topic>Mycology</topic><topic>Photoactivation</topic><topic>Photolysis</topic><topic>Raw sewage</topic><topic>Reduction</topic><topic>Risk assessment</topic><topic>Room temperature</topic><topic>Seawater</topic><topic>Sewage</topic><topic>Viruses</topic><topic>Water analysis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pedrosa de Macena, Lorena da Graça</creatorcontrib><creatorcontrib>Pereira, Joseane Simone de Oliveira</creatorcontrib><creatorcontrib>da Silva, Jansen Couto</creatorcontrib><creatorcontrib>Ferreira, Fernando César</creatorcontrib><creatorcontrib>Maranhão, Adriana Gonçalves</creatorcontrib><creatorcontrib>Lanzarini, Natália Maria</creatorcontrib><creatorcontrib>Miagostovich, Marize Pereira</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Virology and AIDS Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Brazilian journal of microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pedrosa de Macena, Lorena da Graça</au><au>Pereira, Joseane Simone de Oliveira</au><au>da Silva, Jansen Couto</au><au>Ferreira, Fernando César</au><au>Maranhão, Adriana Gonçalves</au><au>Lanzarini, Natália Maria</au><au>Miagostovich, Marize Pereira</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantification of infectious Human mastadenovirus in environmental matrices using PMAxx-qPCR</atitle><jtitle>Brazilian journal of microbiology</jtitle><stitle>Braz J Microbiol</stitle><addtitle>Braz J Microbiol</addtitle><date>2022-09-01</date><risdate>2022</risdate><volume>53</volume><issue>3</issue><spage>1465</spage><epage>1471</epage><pages>1465-1471</pages><issn>1517-8382</issn><eissn>1678-4405</eissn><abstract>Molecular methodologies providing data on viral concentration and infectivity have been successfully used in environmental virology, supporting quantitative risk assessment studies. The present study aimed to assess
human mastadenovirus
(HAdV) intact particles using a derivative of propidium monoazide associated with qPCR (PMAxx-qPCR) in aquatic matrices. Initially, different concentrations of PMAxx were evaluated to establish an optimal protocol for treating different naturally contaminated matrices, using 10 min incubation in the dark at 200 rpm at room temperature and 15 min of photoactivation in the PMA-Lite™ LED photolysis device. There was no significant reduction in the quantification of infectious HAdV with increasing concentration of PMAxx used (20 μM, 50 μM, and 100 μM), except for sewage samples. In this matrix, a reduction of 5.01 log of genomic copies (GC)/L was observed from the concentration of 50 μM and revealed 100% HAdV particles with damaged capsids. On the other hand, the mean reduction of 0.51 log in stool samples using the same concentration mentioned above demonstrated 83% of damaged particles eliminated in the stool. Following, 50 μM PMAxx-qPCR protocol revealed a log reduction of 0.91, 0.67, and 1.05 in other samples of raw sewage, brackish, and seawater where HAdV concentration reached 1.47 × 10
4
, 6.81 × 10
2
, and 2.33 × 10
2
GC/L, respectively. Fifty micrometers of PMAxx protocol helped screen intact viruses from different matrices, including sea and brackish water.</abstract><cop>Cham</cop><pub>Springer International Publishing</pub><pmid>35666431</pmid><doi>10.1007/s42770-022-00775-5</doi><tpages>7</tpages><orcidid>https://orcid.org/0000-0003-1420-6010</orcidid><oa>free_for_read</oa></addata></record> |
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| ispartof | Brazilian journal of microbiology, 2022-09, Vol.53 (3), p.1465-1471 |
| issn | 1517-8382 1678-4405 |
| language | eng |
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| source | Springer Link; PubMed Central |
| subjects | Biomedical and Life Sciences Brackish water Capsids Chemical analysis Environmental Microbiology - Research Paper Food Microbiology Infectivity Life Sciences Medical Microbiology Microbial Ecology Microbial Genetics and Genomics Microbiology Micrometers Mycology Photoactivation Photolysis Raw sewage Reduction Risk assessment Room temperature Seawater Sewage Viruses Water analysis |
| title | Quantification of infectious Human mastadenovirus in environmental matrices using PMAxx-qPCR |
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