HLA-DQA1 allele typing by nonisotopic PCR-LIS-SSCP
In the present study we used a simple and reliable method for HLA-DQA1 allele typing based on the single-stranded conformation polymorphism (SSCP) properties of DNA molecules obtained by PCR. The technique consists of PCR amplification of a DNA fragment comprising the second exon of the HLA-DQA1 gen...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Associação Brasileira de Divulgação Científica
2001-07-01
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| Series: | Brazilian Journal of Medical and Biological Research |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2001000700005 |
| _version_ | 1818061543232765952 |
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| author | M.C. Abba M.A. Gómez C.D. Golijow |
| author_facet | M.C. Abba M.A. Gómez C.D. Golijow |
| author_sort | M.C. Abba |
| collection | DOAJ |
| description | In the present study we used a simple and reliable method for HLA-DQA1 allele typing based on the single-stranded conformation polymorphism (SSCP) properties of DNA molecules obtained by PCR. The technique consists of PCR amplification of a DNA fragment comprising the second exon of the HLA-DQA1 gene, amplicon denaturation using a low ionic strength solution (LIS), and electrophoresis on a small native polyacrylamide gel, followed by a rapid silver staining procedure. In order to validate the technique and to obtain the allele patterns for the DQA1 gene, 50 cervical samples were typed using this methodology and the commercial Amplitype® HLA DQA1 Amplification and Typing kit. All the alleles detected with the kit were characterized by the LIS-SSCP approach. This procedure proved to be useful for population screening and typing of the DQA1 gene as well as for detecting new alleles or mutations in the donor-recipient molecular matching of HLA class II genes. |
| first_indexed | 2024-12-10T13:49:59Z |
| format | Article |
| id | doaj.art-d0a04d85098d4175a0786d372913e8b7 |
| institution | Directory Open Access Journal |
| issn | 0100-879X 1414-431X |
| language | English |
| last_indexed | 2024-12-10T13:49:59Z |
| publishDate | 2001-07-01 |
| publisher | Associação Brasileira de Divulgação Científica |
| record_format | Article |
| series | Brazilian Journal of Medical and Biological Research |
| spelling | doaj.art-d0a04d85098d4175a0786d372913e8b72022-12-22T01:46:16ZengAssociação Brasileira de Divulgação CientíficaBrazilian Journal of Medical and Biological Research0100-879X1414-431X2001-07-0134786786910.1590/S0100-879X2001000700005HLA-DQA1 allele typing by nonisotopic PCR-LIS-SSCPM.C. AbbaM.A. GómezC.D. GolijowIn the present study we used a simple and reliable method for HLA-DQA1 allele typing based on the single-stranded conformation polymorphism (SSCP) properties of DNA molecules obtained by PCR. The technique consists of PCR amplification of a DNA fragment comprising the second exon of the HLA-DQA1 gene, amplicon denaturation using a low ionic strength solution (LIS), and electrophoresis on a small native polyacrylamide gel, followed by a rapid silver staining procedure. In order to validate the technique and to obtain the allele patterns for the DQA1 gene, 50 cervical samples were typed using this methodology and the commercial Amplitype® HLA DQA1 Amplification and Typing kit. All the alleles detected with the kit were characterized by the LIS-SSCP approach. This procedure proved to be useful for population screening and typing of the DQA1 gene as well as for detecting new alleles or mutations in the donor-recipient molecular matching of HLA class II genes.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2001000700005HLA-DQA1PCRLIS-SSCP |
| spellingShingle | M.C. Abba M.A. Gómez C.D. Golijow HLA-DQA1 allele typing by nonisotopic PCR-LIS-SSCP Brazilian Journal of Medical and Biological Research HLA-DQA1 PCR LIS-SSCP |
| title | HLA-DQA1 allele typing by nonisotopic PCR-LIS-SSCP |
| title_full | HLA-DQA1 allele typing by nonisotopic PCR-LIS-SSCP |
| title_fullStr | HLA-DQA1 allele typing by nonisotopic PCR-LIS-SSCP |
| title_full_unstemmed | HLA-DQA1 allele typing by nonisotopic PCR-LIS-SSCP |
| title_short | HLA-DQA1 allele typing by nonisotopic PCR-LIS-SSCP |
| title_sort | hla dqa1 allele typing by nonisotopic pcr lis sscp |
| topic | HLA-DQA1 PCR LIS-SSCP |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2001000700005 |
| work_keys_str_mv | AT mcabba hladqa1alleletypingbynonisotopicpcrlissscp AT magomez hladqa1alleletypingbynonisotopicpcrlissscp AT cdgolijow hladqa1alleletypingbynonisotopicpcrlissscp |