STANDARDIZATION OF PROCEDURES OF Plasmodium falciparum ANTIGEN PREPARATION FOR SEROLOGIC TESTS
The objective of the present study is to standardize the technical variables for preparation and storage of Plasmodium falciparum and of antigen components extracted with the amphoteric detergent Zwittergent. P. falciparum obtained from in vitro culture was stored at different temperatures and for d...
Published in: | Revista do Instituto de Medicina Tropical de São Paulo |
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Language: | English |
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Universidade de São Paulo (USP)
1998
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Online Access: | https://doi.org/10.1590/S0036-46651998000500008 https://doaj.org/article/74e54358da47471288c62ba9a5b459af |
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ftdoajarticles:oai:doaj.org/article:74e54358da47471288c62ba9a5b459af 2024-09-09T19:28:12+00:00 STANDARDIZATION OF PROCEDURES OF Plasmodium falciparum ANTIGEN PREPARATION FOR SEROLOGIC TESTS Sandra L.M. AVILA Tania R. TOZETTO-MENDOZA Viviana G. ARRUK Antonio Walter FERREIRA 1998-09-01T00:00:00Z https://doi.org/10.1590/S0036-46651998000500008 https://doaj.org/article/74e54358da47471288c62ba9a5b459af EN eng Universidade de São Paulo (USP) http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0036-46651998000500008 https://doaj.org/toc/0036-4665 https://doaj.org/toc/1678-9946 doi:10.1590/S0036-46651998000500008 0036-4665 1678-9946 https://doaj.org/article/74e54358da47471288c62ba9a5b459af Revista do Instituto de Medicina Tropical de São Paulo, Vol 40, Iss 5 (1998) Malaria Plasmodium falciparum Antigen ELISA Western blotting Serodiagnosis Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 1998 ftdoajarticles https://doi.org/10.1590/S0036-46651998000500008 2024-08-05T17:49:30Z The objective of the present study is to standardize the technical variables for preparation and storage of Plasmodium falciparum and of antigen components extracted with the amphoteric detergent Zwittergent. P. falciparum obtained from in vitro culture was stored at different temperatures and for different periods of time. For each variable, antigen components of the parasite were extracted in the presence or absence of protease inhibitors and submitted or not to later dialysis. Products were stored for 15, 30 and 60 days at different temperatures and immunological activity of each extract was determined by SDS-PAGE and ELISA using positive or negative standard sera for the presence of IgG directed to blood stage antigens of P. falciparum. Antigen extracts obtained from parasites stored at -20oC up to 10 days or at -70oC for 2 months presented the best results, showing well-defined bands on SDS-PAGE and Western blots and presenting absorbance values in ELISA that permitted safe differentiation between positive and negative sera. O objetivo deste estudo foi padronizar variáveis técnicas para o armazenamento de Plasmodium falciparum e de seus componentes antigênicos. Sedimentos de parasitas foram obtidos do cultivo in vitro de P.falciparum e estocados em diferentes temperaturas por diferentes períodos de tempo. De cada variável, foram extraídos os componentes antigênicos com detergente anfótero Zwittergent na presença e na ausência de inibidores de proteases e submetidos ou não a posterior diálise. Os produtos foram estocados por 15, 30 e 60 dias em diferentes temperaturas e caracterizados por SDS-PAGE. A atividade antigênica de cada extrato foi determinada por ELISA e Western blotting usando soros positivos e negativos para anticorpos IgG anti-formas eritrocitárias de P.falciparum. Os extratos antigênicos obtidos de parasitas estocados até 10 dias a _20ºC ou por 2 meses a _70ºC e tratados com inibidores de proteases, sob as diferentes condições de armazenamento, apresentaram melhor definição das bandas protéicas ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Sob’ ENVELOPE(66.156,66.156,66.322,66.322) Revista do Instituto de Medicina Tropical de São Paulo 40 5 309 316 |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Malaria Plasmodium falciparum Antigen ELISA Western blotting Serodiagnosis Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
Malaria Plasmodium falciparum Antigen ELISA Western blotting Serodiagnosis Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Sandra L.M. AVILA Tania R. TOZETTO-MENDOZA Viviana G. ARRUK Antonio Walter FERREIRA STANDARDIZATION OF PROCEDURES OF Plasmodium falciparum ANTIGEN PREPARATION FOR SEROLOGIC TESTS |
topic_facet |
Malaria Plasmodium falciparum Antigen ELISA Western blotting Serodiagnosis Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
The objective of the present study is to standardize the technical variables for preparation and storage of Plasmodium falciparum and of antigen components extracted with the amphoteric detergent Zwittergent. P. falciparum obtained from in vitro culture was stored at different temperatures and for different periods of time. For each variable, antigen components of the parasite were extracted in the presence or absence of protease inhibitors and submitted or not to later dialysis. Products were stored for 15, 30 and 60 days at different temperatures and immunological activity of each extract was determined by SDS-PAGE and ELISA using positive or negative standard sera for the presence of IgG directed to blood stage antigens of P. falciparum. Antigen extracts obtained from parasites stored at -20oC up to 10 days or at -70oC for 2 months presented the best results, showing well-defined bands on SDS-PAGE and Western blots and presenting absorbance values in ELISA that permitted safe differentiation between positive and negative sera. O objetivo deste estudo foi padronizar variáveis técnicas para o armazenamento de Plasmodium falciparum e de seus componentes antigênicos. Sedimentos de parasitas foram obtidos do cultivo in vitro de P.falciparum e estocados em diferentes temperaturas por diferentes períodos de tempo. De cada variável, foram extraídos os componentes antigênicos com detergente anfótero Zwittergent na presença e na ausência de inibidores de proteases e submetidos ou não a posterior diálise. Os produtos foram estocados por 15, 30 e 60 dias em diferentes temperaturas e caracterizados por SDS-PAGE. A atividade antigênica de cada extrato foi determinada por ELISA e Western blotting usando soros positivos e negativos para anticorpos IgG anti-formas eritrocitárias de P.falciparum. Os extratos antigênicos obtidos de parasitas estocados até 10 dias a _20ºC ou por 2 meses a _70ºC e tratados com inibidores de proteases, sob as diferentes condições de armazenamento, apresentaram melhor definição das bandas protéicas ... |
format |
Article in Journal/Newspaper |
author |
Sandra L.M. AVILA Tania R. TOZETTO-MENDOZA Viviana G. ARRUK Antonio Walter FERREIRA |
author_facet |
Sandra L.M. AVILA Tania R. TOZETTO-MENDOZA Viviana G. ARRUK Antonio Walter FERREIRA |
author_sort |
Sandra L.M. AVILA |
title |
STANDARDIZATION OF PROCEDURES OF Plasmodium falciparum ANTIGEN PREPARATION FOR SEROLOGIC TESTS |
title_short |
STANDARDIZATION OF PROCEDURES OF Plasmodium falciparum ANTIGEN PREPARATION FOR SEROLOGIC TESTS |
title_full |
STANDARDIZATION OF PROCEDURES OF Plasmodium falciparum ANTIGEN PREPARATION FOR SEROLOGIC TESTS |
title_fullStr |
STANDARDIZATION OF PROCEDURES OF Plasmodium falciparum ANTIGEN PREPARATION FOR SEROLOGIC TESTS |
title_full_unstemmed |
STANDARDIZATION OF PROCEDURES OF Plasmodium falciparum ANTIGEN PREPARATION FOR SEROLOGIC TESTS |
title_sort |
standardization of procedures of plasmodium falciparum antigen preparation for serologic tests |
publisher |
Universidade de São Paulo (USP) |
publishDate |
1998 |
url |
https://doi.org/10.1590/S0036-46651998000500008 https://doaj.org/article/74e54358da47471288c62ba9a5b459af |
long_lat |
ENVELOPE(66.156,66.156,66.322,66.322) |
geographic |
Arctic Sob’ |
geographic_facet |
Arctic Sob’ |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Revista do Instituto de Medicina Tropical de São Paulo, Vol 40, Iss 5 (1998) |
op_relation |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0036-46651998000500008 https://doaj.org/toc/0036-4665 https://doaj.org/toc/1678-9946 doi:10.1590/S0036-46651998000500008 0036-4665 1678-9946 https://doaj.org/article/74e54358da47471288c62ba9a5b459af |
op_doi |
https://doi.org/10.1590/S0036-46651998000500008 |
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Revista do Instituto de Medicina Tropical de São Paulo |
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40 |
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