Quantification of Toxoplasma gondii Bradyzoites
A simple 2-step method to selectively quantify functional bradyzoites of Toxoplasma gondii is described. The method was developed to quantify bradyzoites produced in cell cultures that also contain tachyzoites. The selection step comprises incubation of bradyzoites and tachyzoites in 0.026% pepsin a...
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| Veröffentlicht in: | The Journal of parasitology 1996-04, Vol.82 (2), p.330-332 |
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| container_title | The Journal of parasitology |
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| creator | Popiel, I. (Heska Corp., Fort Collins, CO.) Gold, M.C Booth, K.S |
| description | A simple 2-step method to selectively quantify functional bradyzoites of Toxoplasma gondii is described. The method was developed to quantify bradyzoites produced in cell cultures that also contain tachyzoites. The selection step comprises incubation of bradyzoites and tachyzoites in 0.026% pepsin at 37 C for 30 min. This treatment kills all tachyzoites, whereas all bradyzoites survive. A modified plaque assay is then used to quantify the surviving bradyzoites. Plaque assay cultures are scored according to levels of infection that correlate with numbers of bradyzoites added to each well. The assay can detect log differences in the range of 2 x 100-2 x 104bradyzoites per sample. This procedure is simple to perform and provides an efficient way of comparing numbers of functional bradyzoites in multiple samples that also contain tachyzoites. |
| doi_str_mv | 10.2307/3284172 |
| format | Article |
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(Heska Corp., Fort Collins, CO.) ; Gold, M.C ; Booth, K.S</creator><creatorcontrib>Popiel, I. (Heska Corp., Fort Collins, CO.) ; Gold, M.C ; Booth, K.S</creatorcontrib><description>A simple 2-step method to selectively quantify functional bradyzoites of Toxoplasma gondii is described. The method was developed to quantify bradyzoites produced in cell cultures that also contain tachyzoites. The selection step comprises incubation of bradyzoites and tachyzoites in 0.026% pepsin at 37 C for 30 min. This treatment kills all tachyzoites, whereas all bradyzoites survive. A modified plaque assay is then used to quantify the surviving bradyzoites. Plaque assay cultures are scored according to levels of infection that correlate with numbers of bradyzoites added to each well. The assay can detect log differences in the range of 2 x 100-2 x 104bradyzoites per sample. This procedure is simple to perform and provides an efficient way of comparing numbers of functional bradyzoites in multiple samples that also contain tachyzoites.</description><identifier>ISSN: 0022-3395</identifier><identifier>EISSN: 1937-2345</identifier><identifier>DOI: 10.2307/3284172</identifier><identifier>PMID: 8604108</identifier><identifier>CODEN: JOPAA2</identifier><language>eng</language><publisher>Lawrence, KS: American Society of Parasitologists</publisher><subject>Animals ; Biological and medical sciences ; Biological Assay ; Bradyzoites ; Cats ; CELL CULTURE ; Cell culture techniques ; Cells, Cultured ; COUNTING ; CULTIVO DE CELULAS ; CULTURE DE CELLULE ; Cysts ; DEVELOPMENTAL STAGES ; ETAPAS DE DESARROLLO ; Experimental protozoal diseases and models ; Fibroblasts - parasitology ; Gastrointestinal Agents ; Humans ; Immunity ; Infections ; Infectious diseases ; INFECTIVITY ; Medical sciences ; Mice ; Oocysts ; Parasitic diseases ; PATHOGENICITY ; PATOGENICIDAD ; PEPSIN ; Pepsin A ; PEPSINA ; PEPSINE ; Plaque assay ; POUVOIR PATHOGENE ; Protozoal diseases ; Quantification ; Research Notes ; STADE DE DEVELOPPEMENT ; TACHYZOITES ; Toxoplasma - growth & development ; TOXOPLASMA GONDII ; VACCINE DEVELOPMENT ; Viral Plaque Assay</subject><ispartof>The Journal of parasitology, 1996-04, Vol.82 (2), p.330-332</ispartof><rights>Copyright 1996 American Society of Parasitologists</rights><rights>1996 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c387t-48a38948a6bb3629db6c52b35ddedc0fb230891d047dc3ef06c463eb355e10743</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/3284172$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/3284172$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>314,776,780,799,27901,27902,57992,58225</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3049391$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8604108$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Popiel, I. (Heska Corp., Fort Collins, CO.)</creatorcontrib><creatorcontrib>Gold, M.C</creatorcontrib><creatorcontrib>Booth, K.S</creatorcontrib><title>Quantification of Toxoplasma gondii Bradyzoites</title><title>The Journal of parasitology</title><addtitle>J Parasitol</addtitle><description>A simple 2-step method to selectively quantify functional bradyzoites of Toxoplasma gondii is described. The method was developed to quantify bradyzoites produced in cell cultures that also contain tachyzoites. The selection step comprises incubation of bradyzoites and tachyzoites in 0.026% pepsin at 37 C for 30 min. This treatment kills all tachyzoites, whereas all bradyzoites survive. A modified plaque assay is then used to quantify the surviving bradyzoites. Plaque assay cultures are scored according to levels of infection that correlate with numbers of bradyzoites added to each well. The assay can detect log differences in the range of 2 x 100-2 x 104bradyzoites per sample. This procedure is simple to perform and provides an efficient way of comparing numbers of functional bradyzoites in multiple samples that also contain tachyzoites.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Biological Assay</subject><subject>Bradyzoites</subject><subject>Cats</subject><subject>CELL CULTURE</subject><subject>Cell culture techniques</subject><subject>Cells, Cultured</subject><subject>COUNTING</subject><subject>CULTIVO DE CELULAS</subject><subject>CULTURE DE CELLULE</subject><subject>Cysts</subject><subject>DEVELOPMENTAL STAGES</subject><subject>ETAPAS DE DESARROLLO</subject><subject>Experimental protozoal diseases and models</subject><subject>Fibroblasts - parasitology</subject><subject>Gastrointestinal Agents</subject><subject>Humans</subject><subject>Immunity</subject><subject>Infections</subject><subject>Infectious diseases</subject><subject>INFECTIVITY</subject><subject>Medical sciences</subject><subject>Mice</subject><subject>Oocysts</subject><subject>Parasitic diseases</subject><subject>PATHOGENICITY</subject><subject>PATOGENICIDAD</subject><subject>PEPSIN</subject><subject>Pepsin A</subject><subject>PEPSINA</subject><subject>PEPSINE</subject><subject>Plaque assay</subject><subject>POUVOIR PATHOGENE</subject><subject>Protozoal diseases</subject><subject>Quantification</subject><subject>Research Notes</subject><subject>STADE DE DEVELOPPEMENT</subject><subject>TACHYZOITES</subject><subject>Toxoplasma - growth & development</subject><subject>TOXOPLASMA GONDII</subject><subject>VACCINE DEVELOPMENT</subject><subject>Viral Plaque Assay</subject><issn>0022-3395</issn><issn>1937-2345</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1LxDAQhoMo67qKv0DoQfRUnXSaJjmq-AWCiLvnkiapRNpmTVpw_fV2sejRw8wc5uGd4SHkmMJFhsAvMRM55dkOmVOJPM0wZ7tkDpBlKaJk--QgxncAYGPNyEwUkFMQc3L5Mqiud7XTqne-S3ydLP2nXzcqtip5851xLrkOymy-vOttPCR7tWqiPZrmgqzubpc3D-nT8_3jzdVTqlHwPs2FQiHHXlQVFpk0VaFZViEzxhoNdTU-LSQ1kHOj0dZQ6LxAOwLMUuA5LsjZT-46-I_Bxr5sXdS2aVRn_RBLzqUQosB_QcoKBoxtwfMfUAcfY7B1uQ6uVWFTUii3DsvJ4UieTJFD1Vrzy03Sxv3ptFdRq6YOqtMu_mIIuURJ_7D32Pvw_7Va-VK9hTFp9Uql5MDoqAi_AZsMilY</recordid><startdate>19960401</startdate><enddate>19960401</enddate><creator>Popiel, I. (Heska Corp., Fort Collins, CO.)</creator><creator>Gold, M.C</creator><creator>Booth, K.S</creator><general>American Society of Parasitologists</general><scope>FBQ</scope><scope>IQODW</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>M7N</scope><scope>7X8</scope></search><sort><creationdate>19960401</creationdate><title>Quantification of Toxoplasma gondii Bradyzoites</title><author>Popiel, I. (Heska Corp., Fort Collins, CO.) ; Gold, M.C ; Booth, K.S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c387t-48a38948a6bb3629db6c52b35ddedc0fb230891d047dc3ef06c463eb355e10743</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Biological Assay</topic><topic>Bradyzoites</topic><topic>Cats</topic><topic>CELL CULTURE</topic><topic>Cell culture techniques</topic><topic>Cells, Cultured</topic><topic>COUNTING</topic><topic>CULTIVO DE CELULAS</topic><topic>CULTURE DE CELLULE</topic><topic>Cysts</topic><topic>DEVELOPMENTAL STAGES</topic><topic>ETAPAS DE DESARROLLO</topic><topic>Experimental protozoal diseases and models</topic><topic>Fibroblasts - parasitology</topic><topic>Gastrointestinal Agents</topic><topic>Humans</topic><topic>Immunity</topic><topic>Infections</topic><topic>Infectious diseases</topic><topic>INFECTIVITY</topic><topic>Medical sciences</topic><topic>Mice</topic><topic>Oocysts</topic><topic>Parasitic diseases</topic><topic>PATHOGENICITY</topic><topic>PATOGENICIDAD</topic><topic>PEPSIN</topic><topic>Pepsin A</topic><topic>PEPSINA</topic><topic>PEPSINE</topic><topic>Plaque assay</topic><topic>POUVOIR PATHOGENE</topic><topic>Protozoal diseases</topic><topic>Quantification</topic><topic>Research Notes</topic><topic>STADE DE DEVELOPPEMENT</topic><topic>TACHYZOITES</topic><topic>Toxoplasma - growth & development</topic><topic>TOXOPLASMA GONDII</topic><topic>VACCINE DEVELOPMENT</topic><topic>Viral Plaque Assay</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Popiel, I. (Heska Corp., Fort Collins, CO.)</creatorcontrib><creatorcontrib>Gold, M.C</creatorcontrib><creatorcontrib>Booth, K.S</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of parasitology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Popiel, I. (Heska Corp., Fort Collins, CO.)</au><au>Gold, M.C</au><au>Booth, K.S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantification of Toxoplasma gondii Bradyzoites</atitle><jtitle>The Journal of parasitology</jtitle><addtitle>J Parasitol</addtitle><date>1996-04-01</date><risdate>1996</risdate><volume>82</volume><issue>2</issue><spage>330</spage><epage>332</epage><pages>330-332</pages><issn>0022-3395</issn><eissn>1937-2345</eissn><coden>JOPAA2</coden><abstract>A simple 2-step method to selectively quantify functional bradyzoites of Toxoplasma gondii is described. The method was developed to quantify bradyzoites produced in cell cultures that also contain tachyzoites. The selection step comprises incubation of bradyzoites and tachyzoites in 0.026% pepsin at 37 C for 30 min. This treatment kills all tachyzoites, whereas all bradyzoites survive. A modified plaque assay is then used to quantify the surviving bradyzoites. Plaque assay cultures are scored according to levels of infection that correlate with numbers of bradyzoites added to each well. The assay can detect log differences in the range of 2 x 100-2 x 104bradyzoites per sample. This procedure is simple to perform and provides an efficient way of comparing numbers of functional bradyzoites in multiple samples that also contain tachyzoites.</abstract><cop>Lawrence, KS</cop><pub>American Society of Parasitologists</pub><pmid>8604108</pmid><doi>10.2307/3284172</doi><tpages>3</tpages></addata></record> |
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| language | eng |
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| source | Jstor Complete Legacy; MEDLINE |
| subjects | Animals Biological and medical sciences Biological Assay Bradyzoites Cats CELL CULTURE Cell culture techniques Cells, Cultured COUNTING CULTIVO DE CELULAS CULTURE DE CELLULE Cysts DEVELOPMENTAL STAGES ETAPAS DE DESARROLLO Experimental protozoal diseases and models Fibroblasts - parasitology Gastrointestinal Agents Humans Immunity Infections Infectious diseases INFECTIVITY Medical sciences Mice Oocysts Parasitic diseases PATHOGENICITY PATOGENICIDAD PEPSIN Pepsin A PEPSINA PEPSINE Plaque assay POUVOIR PATHOGENE Protozoal diseases Quantification Research Notes STADE DE DEVELOPPEMENT TACHYZOITES Toxoplasma - growth & development TOXOPLASMA GONDII VACCINE DEVELOPMENT Viral Plaque Assay |
| title | Quantification of Toxoplasma gondii Bradyzoites |
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