Standardization of Semi-Quantitative Dot Blotting Assay—Application in the Diagnosis, Follow-Up, and Relapse of Paracoccidioidomycosis
Introduction: This study standardized a semi-quantitative dot blotting assay (DB) and a quantitative real-time polymerase chain reaction (qPCR) to detect specific antibodies for <i>Paracoccidioides brasiliensis</i> and its DNA in PCM patients. Methodology: We evaluated 42 confirmed PCM p...
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MDPI AG
2024-02-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/12/2/351 |
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| author | Beatriz Aparecida Soares Pereira Ricardo de Souza Cavalcante Vera Lucia Pereira-Chioccola Marcia de Souza Carvalho Melhem Lídia Raquel de Carvalho Rinaldo Poncio Mendes |
| author_facet | Beatriz Aparecida Soares Pereira Ricardo de Souza Cavalcante Vera Lucia Pereira-Chioccola Marcia de Souza Carvalho Melhem Lídia Raquel de Carvalho Rinaldo Poncio Mendes |
| author_sort | Beatriz Aparecida Soares Pereira |
| collection | DOAJ |
| description | Introduction: This study standardized a semi-quantitative dot blotting assay (DB) and a quantitative real-time polymerase chain reaction (qPCR) to detect specific antibodies for <i>Paracoccidioides brasiliensis</i> and its DNA in PCM patients. Methodology: We evaluated 42 confirmed PCM patients upon admission using a serological double agar gel immunodiffusion test (DID), DB, and molecular tests (qPCR in total blood). The control groups included 42 healthy individuals and 37 patients with other infectious diseases. The serological progress during treatment was evaluated in eight patients, and there was a relapse diagnosis in ten patients using the Pb B.339 strain antigen. The cut-off points for the serological tests were determined by a receiver operator characteristic curve. Results: The DB and DID tests showed similar accuracy, but the DB identified lower antibody concentrations. Cross-reactions were absent in the DB assay. In the relapse diagnoses, DB exhibited much higher sensitivity (90%) than DID (30%). Conclusions: A DB assay is easier and faster than a DID test to be performed; DB and DID tests show the same accuracy, while blood qPCR is not recommended in the diagnosis at the time of admission; cross-reactions were not observed with other systemic diseases; DB and DID tests are useful for treatment monitoring PCM patients; and a DB assay is the choice for diagnosing relapse. These findings support the introduction of semi-quantitative DB assays in clinical laboratories. |
| first_indexed | 2024-03-07T22:21:12Z |
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| id | doaj.art-b960e39a5acd4e289af7715f96743d8c |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-07T22:21:12Z |
| publishDate | 2024-02-01 |
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| series | Microorganisms |
| spelling | doaj.art-b960e39a5acd4e289af7715f96743d8c2024-02-23T15:28:18ZengMDPI AGMicroorganisms2076-26072024-02-0112235110.3390/microorganisms12020351Standardization of Semi-Quantitative Dot Blotting Assay—Application in the Diagnosis, Follow-Up, and Relapse of ParacoccidioidomycosisBeatriz Aparecida Soares Pereira0Ricardo de Souza Cavalcante1Vera Lucia Pereira-Chioccola2Marcia de Souza Carvalho Melhem3Lídia Raquel de Carvalho4Rinaldo Poncio Mendes5UNESP Botucatu, School of Medicine—Discipline of Infectology, São Paulo State University, Botucatu 18618-689, São Paulo State, BrazilUNESP Botucatu, School of Medicine—Discipline of Infectology, São Paulo State University, Botucatu 18618-689, São Paulo State, BrazilAdolfo Lutz Institute, São Paulo 01246-000, São Paulo State, BrazilUNESP Botucatu, School of Medicine—Discipline of Infectology, São Paulo State University, Botucatu 18618-689, São Paulo State, BrazilInstitute of Biosciences—Department of Biodiversity and Biostatistics, UNESP Botucatu, São Paulo State University, Botucatu 18618-689, São Paulo State, BrazilUNESP Botucatu, School of Medicine—Discipline of Infectology, São Paulo State University, Botucatu 18618-689, São Paulo State, BrazilIntroduction: This study standardized a semi-quantitative dot blotting assay (DB) and a quantitative real-time polymerase chain reaction (qPCR) to detect specific antibodies for <i>Paracoccidioides brasiliensis</i> and its DNA in PCM patients. Methodology: We evaluated 42 confirmed PCM patients upon admission using a serological double agar gel immunodiffusion test (DID), DB, and molecular tests (qPCR in total blood). The control groups included 42 healthy individuals and 37 patients with other infectious diseases. The serological progress during treatment was evaluated in eight patients, and there was a relapse diagnosis in ten patients using the Pb B.339 strain antigen. The cut-off points for the serological tests were determined by a receiver operator characteristic curve. Results: The DB and DID tests showed similar accuracy, but the DB identified lower antibody concentrations. Cross-reactions were absent in the DB assay. In the relapse diagnoses, DB exhibited much higher sensitivity (90%) than DID (30%). Conclusions: A DB assay is easier and faster than a DID test to be performed; DB and DID tests show the same accuracy, while blood qPCR is not recommended in the diagnosis at the time of admission; cross-reactions were not observed with other systemic diseases; DB and DID tests are useful for treatment monitoring PCM patients; and a DB assay is the choice for diagnosing relapse. These findings support the introduction of semi-quantitative DB assays in clinical laboratories.https://www.mdpi.com/2076-2607/12/2/351paracoccidioidomycosisdot blottingdiagnosisserologyrelapse |
| spellingShingle | Beatriz Aparecida Soares Pereira Ricardo de Souza Cavalcante Vera Lucia Pereira-Chioccola Marcia de Souza Carvalho Melhem Lídia Raquel de Carvalho Rinaldo Poncio Mendes Standardization of Semi-Quantitative Dot Blotting Assay—Application in the Diagnosis, Follow-Up, and Relapse of Paracoccidioidomycosis Microorganisms paracoccidioidomycosis dot blotting diagnosis serology relapse |
| title | Standardization of Semi-Quantitative Dot Blotting Assay—Application in the Diagnosis, Follow-Up, and Relapse of Paracoccidioidomycosis |
| title_full | Standardization of Semi-Quantitative Dot Blotting Assay—Application in the Diagnosis, Follow-Up, and Relapse of Paracoccidioidomycosis |
| title_fullStr | Standardization of Semi-Quantitative Dot Blotting Assay—Application in the Diagnosis, Follow-Up, and Relapse of Paracoccidioidomycosis |
| title_full_unstemmed | Standardization of Semi-Quantitative Dot Blotting Assay—Application in the Diagnosis, Follow-Up, and Relapse of Paracoccidioidomycosis |
| title_short | Standardization of Semi-Quantitative Dot Blotting Assay—Application in the Diagnosis, Follow-Up, and Relapse of Paracoccidioidomycosis |
| title_sort | standardization of semi quantitative dot blotting assay application in the diagnosis follow up and relapse of paracoccidioidomycosis |
| topic | paracoccidioidomycosis dot blotting diagnosis serology relapse |
| url | https://www.mdpi.com/2076-2607/12/2/351 |
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