Evaluation of a flow cytometric test for G6PD‐deficient erythrocytes
Objective Glucose‐6‐phosphate dehydrogenase (G6PD) deficiency, an X‐linked recessive disorder, is the commonest erythrocytic enzymopathy worldwide. Reliable diagnosis and severity prediction in G6PD‐deficient/heterozygous females remain challenging. A recently developed flow cytometric test for G6PD...
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| Veröffentlicht in: | Tropical medicine & international health 2021-04, Vol.26 (4), p.462-468 |
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| creator | Kapadia, Alpeshkumar Bipinbhai Sharma, Prashant Jain, Karuna Sachdeva, Man Updesh Singh Bose, Parveen Lata Gupta, Minakshi Khadwal, Alka Rani Bal, Amanjit Das, Reena Varma, Neelam |
| description | Objective
Glucose‐6‐phosphate dehydrogenase (G6PD) deficiency, an X‐linked recessive disorder, is the commonest erythrocytic enzymopathy worldwide. Reliable diagnosis and severity prediction in G6PD‐deficient/heterozygous females remain challenging. A recently developed flow cytometric test for G6PD deficiency has shown promise in precisely identifying deficient females. This paper presents our experiences with this test in a subtropical setting and presents a modification in flow cytometric data acquisition strategy.
Methods
The methaemoglobin reduction + ferryl Hb generation‐based flow cytometric G6PD test was compared with the screening methaemoglobin reduction test (MRT) and confirmatory G6PD enzyme activity assay (EAA) in 20 G6PD‐deficient males, 22 G6PD‐heterozygous/deficient females and 20 controls. Stained cells were also assessed for bright/dim G6PD activity under a fluorescent microscope.
Results
Flow cytometry separated and quantified %bright cells in heterozygous/deficient females, objectively classifying them into 6 normal (>85% bright cells), 14 intermediate (10‐85%) and two G6PD‐deficient ( |
| doi_str_mv | 10.1111/tmi.13547 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_2476566111</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2476566111</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3487-839c2ddc24d70bcb3b627ae4a1d742ba9cdcd98d9789bf176965425227d0c7643</originalsourceid><addsrcrecordid>eNp1kD1OxDAQRi0EYmGh4AIoEg0UWfzvpEQLLEggKJbacmxHBCUx2AmrdByBM3ISDFkokJhmpnj69M0D4ADBGYpz2jXVDBFGxQbYQYSzlCDGN79vmGIs-ATshvAEIaSU8W0wIYQiJvJsB1xevKq6V13l2sSViUrK2q0SPXSusZ2vdNLZ0CWl88mC359_vL0bW1a6sm2XWD90j95F1oY9sFWqOtj99Z6Ch8uL5fwqvblbXM_PblJNaCbSjOQaG6MxNQIWuiAFx0JZqpARFBcq10abPDO5yPKiRILnnFHM4gsGasEpmYLjMffZu5c-VpNNFbSta9Va1weJqeCM8yglokd_0CfX-za2k5hBITKBBYvUyUhp70LwtpTPvmqUHySC8kuujHLlt9zIHq4T-6Kx5pf8sRmB0xFYVbUd_k-Sy9vrMfITxVGDDw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2507787275</pqid></control><display><type>article</type><title>Evaluation of a flow cytometric test for G6PD‐deficient erythrocytes</title><source>Wiley Free Content</source><source>Wiley Online Library Journals Frontfile Complete</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><creator>Kapadia, Alpeshkumar Bipinbhai ; Sharma, Prashant ; Jain, Karuna ; Sachdeva, Man Updesh Singh ; Bose, Parveen Lata ; Gupta, Minakshi ; Khadwal, Alka Rani ; Bal, Amanjit ; Das, Reena ; Varma, Neelam</creator><creatorcontrib>Kapadia, Alpeshkumar Bipinbhai ; Sharma, Prashant ; Jain, Karuna ; Sachdeva, Man Updesh Singh ; Bose, Parveen Lata ; Gupta, Minakshi ; Khadwal, Alka Rani ; Bal, Amanjit ; Das, Reena ; Varma, Neelam</creatorcontrib><description>Objective
Glucose‐6‐phosphate dehydrogenase (G6PD) deficiency, an X‐linked recessive disorder, is the commonest erythrocytic enzymopathy worldwide. Reliable diagnosis and severity prediction in G6PD‐deficient/heterozygous females remain challenging. A recently developed flow cytometric test for G6PD deficiency has shown promise in precisely identifying deficient females. This paper presents our experiences with this test in a subtropical setting and presents a modification in flow cytometric data acquisition strategy.
Methods
The methaemoglobin reduction + ferryl Hb generation‐based flow cytometric G6PD test was compared with the screening methaemoglobin reduction test (MRT) and confirmatory G6PD enzyme activity assay (EAA) in 20 G6PD‐deficient males, 22 G6PD‐heterozygous/deficient females and 20 controls. Stained cells were also assessed for bright/dim G6PD activity under a fluorescent microscope.
Results
Flow cytometry separated and quantified %bright cells in heterozygous/deficient females, objectively classifying them into 6 normal (>85% bright cells), 14 intermediate (10‐85%) and two G6PD‐deficient (<10% bright cells). Concordance with MRT was 89% (55/62 cases) and with EAA was 77% (48/62 cases). Fluorometrically predicted violet laser excitation (405‐nm) with signal acquisition in the 425–475 nm region was a technical advancement noted for the first time in this paper.
Conclusion
Flow cytometry/fluorescence microscopy represent technically straightforward methods for the detection and quantification of G6PD‐deficient erythrocytes. Based on our results, we recommend their application as a first‐line investigation to screen females who are prescribed an oxidant drug like primaquine or dapsone.</description><identifier>ISSN: 1360-2276</identifier><identifier>EISSN: 1365-3156</identifier><identifier>DOI: 10.1111/tmi.13547</identifier><identifier>PMID: 33415798</identifier><language>eng</language><publisher>England: Blackwell Publishing Ltd</publisher><subject>Dapsone ; Data acquisition ; Diaminodiphenylsulfone ; Enzymatic activity ; Enzyme activity ; Erythrocytes ; Females ; Flow cytometry ; Fluorescence ; Fluorescence microscopy ; G6PD ; Glucose 6 phosphate dehydrogenase ; Glucosephosphate dehydrogenase ; Hereditary diseases ; heterozygous female carriers ; laboratory diagnosis ; Oxidants ; Oxidizing agents ; Primaquine ; Reduction</subject><ispartof>Tropical medicine & international health, 2021-04, Vol.26 (4), p.462-468</ispartof><rights>2021 John Wiley & Sons Ltd</rights><rights>2021 John Wiley & Sons Ltd.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c3487-839c2ddc24d70bcb3b627ae4a1d742ba9cdcd98d9789bf176965425227d0c7643</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Ftmi.13547$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Ftmi.13547$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,1427,27901,27902,45550,45551,46384,46808</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33415798$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kapadia, Alpeshkumar Bipinbhai</creatorcontrib><creatorcontrib>Sharma, Prashant</creatorcontrib><creatorcontrib>Jain, Karuna</creatorcontrib><creatorcontrib>Sachdeva, Man Updesh Singh</creatorcontrib><creatorcontrib>Bose, Parveen Lata</creatorcontrib><creatorcontrib>Gupta, Minakshi</creatorcontrib><creatorcontrib>Khadwal, Alka Rani</creatorcontrib><creatorcontrib>Bal, Amanjit</creatorcontrib><creatorcontrib>Das, Reena</creatorcontrib><creatorcontrib>Varma, Neelam</creatorcontrib><title>Evaluation of a flow cytometric test for G6PD‐deficient erythrocytes</title><title>Tropical medicine & international health</title><addtitle>Trop Med Int Health</addtitle><description>Objective
Glucose‐6‐phosphate dehydrogenase (G6PD) deficiency, an X‐linked recessive disorder, is the commonest erythrocytic enzymopathy worldwide. Reliable diagnosis and severity prediction in G6PD‐deficient/heterozygous females remain challenging. A recently developed flow cytometric test for G6PD deficiency has shown promise in precisely identifying deficient females. This paper presents our experiences with this test in a subtropical setting and presents a modification in flow cytometric data acquisition strategy.
Methods
The methaemoglobin reduction + ferryl Hb generation‐based flow cytometric G6PD test was compared with the screening methaemoglobin reduction test (MRT) and confirmatory G6PD enzyme activity assay (EAA) in 20 G6PD‐deficient males, 22 G6PD‐heterozygous/deficient females and 20 controls. Stained cells were also assessed for bright/dim G6PD activity under a fluorescent microscope.
Results
Flow cytometry separated and quantified %bright cells in heterozygous/deficient females, objectively classifying them into 6 normal (>85% bright cells), 14 intermediate (10‐85%) and two G6PD‐deficient (<10% bright cells). Concordance with MRT was 89% (55/62 cases) and with EAA was 77% (48/62 cases). Fluorometrically predicted violet laser excitation (405‐nm) with signal acquisition in the 425–475 nm region was a technical advancement noted for the first time in this paper.
Conclusion
Flow cytometry/fluorescence microscopy represent technically straightforward methods for the detection and quantification of G6PD‐deficient erythrocytes. Based on our results, we recommend their application as a first‐line investigation to screen females who are prescribed an oxidant drug like primaquine or dapsone.</description><subject>Dapsone</subject><subject>Data acquisition</subject><subject>Diaminodiphenylsulfone</subject><subject>Enzymatic activity</subject><subject>Enzyme activity</subject><subject>Erythrocytes</subject><subject>Females</subject><subject>Flow cytometry</subject><subject>Fluorescence</subject><subject>Fluorescence microscopy</subject><subject>G6PD</subject><subject>Glucose 6 phosphate dehydrogenase</subject><subject>Glucosephosphate dehydrogenase</subject><subject>Hereditary diseases</subject><subject>heterozygous female carriers</subject><subject>laboratory diagnosis</subject><subject>Oxidants</subject><subject>Oxidizing agents</subject><subject>Primaquine</subject><subject>Reduction</subject><issn>1360-2276</issn><issn>1365-3156</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><recordid>eNp1kD1OxDAQRi0EYmGh4AIoEg0UWfzvpEQLLEggKJbacmxHBCUx2AmrdByBM3ISDFkokJhmpnj69M0D4ADBGYpz2jXVDBFGxQbYQYSzlCDGN79vmGIs-ATshvAEIaSU8W0wIYQiJvJsB1xevKq6V13l2sSViUrK2q0SPXSusZ2vdNLZ0CWl88mC359_vL0bW1a6sm2XWD90j95F1oY9sFWqOtj99Z6Ch8uL5fwqvblbXM_PblJNaCbSjOQaG6MxNQIWuiAFx0JZqpARFBcq10abPDO5yPKiRILnnFHM4gsGasEpmYLjMffZu5c-VpNNFbSta9Va1weJqeCM8yglokd_0CfX-za2k5hBITKBBYvUyUhp70LwtpTPvmqUHySC8kuujHLlt9zIHq4T-6Kx5pf8sRmB0xFYVbUd_k-Sy9vrMfITxVGDDw</recordid><startdate>202104</startdate><enddate>202104</enddate><creator>Kapadia, Alpeshkumar Bipinbhai</creator><creator>Sharma, Prashant</creator><creator>Jain, Karuna</creator><creator>Sachdeva, Man Updesh Singh</creator><creator>Bose, Parveen Lata</creator><creator>Gupta, Minakshi</creator><creator>Khadwal, Alka Rani</creator><creator>Bal, Amanjit</creator><creator>Das, Reena</creator><creator>Varma, Neelam</creator><general>Blackwell Publishing Ltd</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7T2</scope><scope>7U9</scope><scope>C1K</scope><scope>H94</scope><scope>K9.</scope><scope>M7N</scope><scope>7X8</scope></search><sort><creationdate>202104</creationdate><title>Evaluation of a flow cytometric test for G6PD‐deficient erythrocytes</title><author>Kapadia, Alpeshkumar Bipinbhai ; Sharma, Prashant ; Jain, Karuna ; Sachdeva, Man Updesh Singh ; Bose, Parveen Lata ; Gupta, Minakshi ; Khadwal, Alka Rani ; Bal, Amanjit ; Das, Reena ; Varma, Neelam</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3487-839c2ddc24d70bcb3b627ae4a1d742ba9cdcd98d9789bf176965425227d0c7643</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Dapsone</topic><topic>Data acquisition</topic><topic>Diaminodiphenylsulfone</topic><topic>Enzymatic activity</topic><topic>Enzyme activity</topic><topic>Erythrocytes</topic><topic>Females</topic><topic>Flow cytometry</topic><topic>Fluorescence</topic><topic>Fluorescence microscopy</topic><topic>G6PD</topic><topic>Glucose 6 phosphate dehydrogenase</topic><topic>Glucosephosphate dehydrogenase</topic><topic>Hereditary diseases</topic><topic>heterozygous female carriers</topic><topic>laboratory diagnosis</topic><topic>Oxidants</topic><topic>Oxidizing agents</topic><topic>Primaquine</topic><topic>Reduction</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kapadia, Alpeshkumar Bipinbhai</creatorcontrib><creatorcontrib>Sharma, Prashant</creatorcontrib><creatorcontrib>Jain, Karuna</creatorcontrib><creatorcontrib>Sachdeva, Man Updesh Singh</creatorcontrib><creatorcontrib>Bose, Parveen Lata</creatorcontrib><creatorcontrib>Gupta, Minakshi</creatorcontrib><creatorcontrib>Khadwal, Alka Rani</creatorcontrib><creatorcontrib>Bal, Amanjit</creatorcontrib><creatorcontrib>Das, Reena</creatorcontrib><creatorcontrib>Varma, Neelam</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Health and Safety Science Abstracts (Full archive)</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><jtitle>Tropical medicine & international health</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kapadia, Alpeshkumar Bipinbhai</au><au>Sharma, Prashant</au><au>Jain, Karuna</au><au>Sachdeva, Man Updesh Singh</au><au>Bose, Parveen Lata</au><au>Gupta, Minakshi</au><au>Khadwal, Alka Rani</au><au>Bal, Amanjit</au><au>Das, Reena</au><au>Varma, Neelam</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Evaluation of a flow cytometric test for G6PD‐deficient erythrocytes</atitle><jtitle>Tropical medicine & international health</jtitle><addtitle>Trop Med Int Health</addtitle><date>2021-04</date><risdate>2021</risdate><volume>26</volume><issue>4</issue><spage>462</spage><epage>468</epage><pages>462-468</pages><issn>1360-2276</issn><eissn>1365-3156</eissn><abstract>Objective
Glucose‐6‐phosphate dehydrogenase (G6PD) deficiency, an X‐linked recessive disorder, is the commonest erythrocytic enzymopathy worldwide. Reliable diagnosis and severity prediction in G6PD‐deficient/heterozygous females remain challenging. A recently developed flow cytometric test for G6PD deficiency has shown promise in precisely identifying deficient females. This paper presents our experiences with this test in a subtropical setting and presents a modification in flow cytometric data acquisition strategy.
Methods
The methaemoglobin reduction + ferryl Hb generation‐based flow cytometric G6PD test was compared with the screening methaemoglobin reduction test (MRT) and confirmatory G6PD enzyme activity assay (EAA) in 20 G6PD‐deficient males, 22 G6PD‐heterozygous/deficient females and 20 controls. Stained cells were also assessed for bright/dim G6PD activity under a fluorescent microscope.
Results
Flow cytometry separated and quantified %bright cells in heterozygous/deficient females, objectively classifying them into 6 normal (>85% bright cells), 14 intermediate (10‐85%) and two G6PD‐deficient (<10% bright cells). Concordance with MRT was 89% (55/62 cases) and with EAA was 77% (48/62 cases). Fluorometrically predicted violet laser excitation (405‐nm) with signal acquisition in the 425–475 nm region was a technical advancement noted for the first time in this paper.
Conclusion
Flow cytometry/fluorescence microscopy represent technically straightforward methods for the detection and quantification of G6PD‐deficient erythrocytes. Based on our results, we recommend their application as a first‐line investigation to screen females who are prescribed an oxidant drug like primaquine or dapsone.</abstract><cop>England</cop><pub>Blackwell Publishing Ltd</pub><pmid>33415798</pmid><doi>10.1111/tmi.13547</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Dapsone Data acquisition Diaminodiphenylsulfone Enzymatic activity Enzyme activity Erythrocytes Females Flow cytometry Fluorescence Fluorescence microscopy G6PD Glucose 6 phosphate dehydrogenase Glucosephosphate dehydrogenase Hereditary diseases heterozygous female carriers laboratory diagnosis Oxidants Oxidizing agents Primaquine Reduction |
| title | Evaluation of a flow cytometric test for G6PD‐deficient erythrocytes |
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