Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale
Malaria diagnoses has traditionally been made using thick blood smears, but more sensitive and faster techniques are required to process large numbers of samples in clinical and epidemiological studies and in blood donor screening. Here, we evaluated molecular and serological tools to build a screen...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Fundação Oswaldo Cruz (FIOCRUZ)
2011-09-01
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| Series: | Memorias do Instituto Oswaldo Cruz |
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| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762011000600008&lng=en&tlng=en |
| _version_ | 1797757394791432192 |
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| author | Giselle FMC Lima José E Levi Marcelo P Geraldi Maria Carmen A Sanchez Aluísio AC Segurado Angélica D Hristov Juliana Inoue Maria de Jesus Costa-Nascimento Silvia M Di Santi |
| author_facet | Giselle FMC Lima José E Levi Marcelo P Geraldi Maria Carmen A Sanchez Aluísio AC Segurado Angélica D Hristov Juliana Inoue Maria de Jesus Costa-Nascimento Silvia M Di Santi |
| author_sort | Giselle FMC Lima |
| collection | DOAJ |
| description | Malaria diagnoses has traditionally been made using thick blood smears, but more sensitive and faster techniques are required to process large numbers of samples in clinical and epidemiological studies and in blood donor screening. Here, we evaluated molecular and serological tools to build a screening platform for pooled samples aimed at reducing both the time and the cost of these diagnoses. Positive and negative samples were analysed in individual and pooled experiments using real-time polymerase chain reaction (PCR), nested PCR and an immunochromatographic test. For the individual tests, 46/49 samples were positive by real-time PCR, 46/49 were positive by nested PCR and 32/46 were positive by immunochromatographic test. For the assays performed using pooled samples, 13/15 samples were positive by real-time PCR and nested PCR and 11/15 were positive by immunochromatographic test. These molecular methods demonstrated sensitivity and specificity for both the individual and pooled samples. Due to the advantages of the real-time PCR, such as the fast processing and the closed system, this method should be indicated as the first choice for use in large-scale diagnosis and the nested PCR should be used for species differentiation. However, additional field isolates should be tested to confirm the results achieved using cultured parasites and the serological test should only be adopted as a complementary method for malaria diagnosis. |
| first_indexed | 2024-03-12T18:15:11Z |
| format | Article |
| id | doaj.art-59c449113a834867b23ea9b075c71a1e |
| institution | Directory Open Access Journal |
| issn | 1678-8060 |
| language | English |
| last_indexed | 2024-03-12T18:15:11Z |
| publishDate | 2011-09-01 |
| publisher | Fundação Oswaldo Cruz (FIOCRUZ) |
| record_format | Article |
| series | Memorias do Instituto Oswaldo Cruz |
| spelling | doaj.art-59c449113a834867b23ea9b075c71a1e2023-08-02T09:10:45ZengFundação Oswaldo Cruz (FIOCRUZ)Memorias do Instituto Oswaldo Cruz1678-80602011-09-01106669170010.1590/S0074-02762011000600008S0074-02762011000600008Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scaleGiselle FMC Lima0José E Levi1Marcelo P Geraldi2Maria Carmen A Sanchez3Aluísio AC Segurado4Angélica D Hristov5Juliana Inoue6Maria de Jesus Costa-Nascimento7Silvia M Di Santi8Secretaria da Saúde do Estado de São PauloHospital Sírio-LibanêsLaboratório de VirologiaUniversidade de São PauloUniversidade de São PauloSecretaria da Saúde do Estado de São PauloSecretaria da Saúde do Estado de São PauloSecretaria da Saúde do Estado de São PauloSecretaria da Saúde do Estado de São PauloMalaria diagnoses has traditionally been made using thick blood smears, but more sensitive and faster techniques are required to process large numbers of samples in clinical and epidemiological studies and in blood donor screening. Here, we evaluated molecular and serological tools to build a screening platform for pooled samples aimed at reducing both the time and the cost of these diagnoses. Positive and negative samples were analysed in individual and pooled experiments using real-time polymerase chain reaction (PCR), nested PCR and an immunochromatographic test. For the individual tests, 46/49 samples were positive by real-time PCR, 46/49 were positive by nested PCR and 32/46 were positive by immunochromatographic test. For the assays performed using pooled samples, 13/15 samples were positive by real-time PCR and nested PCR and 11/15 were positive by immunochromatographic test. These molecular methods demonstrated sensitivity and specificity for both the individual and pooled samples. Due to the advantages of the real-time PCR, such as the fast processing and the closed system, this method should be indicated as the first choice for use in large-scale diagnosis and the nested PCR should be used for species differentiation. However, additional field isolates should be tested to confirm the results achieved using cultured parasites and the serological test should only be adopted as a complementary method for malaria diagnosis.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762011000600008&lng=en&tlng=enmalariareal-time PCRnested PCRPlasmodiumdiagnosis |
| spellingShingle | Giselle FMC Lima José E Levi Marcelo P Geraldi Maria Carmen A Sanchez Aluísio AC Segurado Angélica D Hristov Juliana Inoue Maria de Jesus Costa-Nascimento Silvia M Di Santi Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale Memorias do Instituto Oswaldo Cruz malaria real-time PCR nested PCR Plasmodium diagnosis |
| title | Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale |
| title_full | Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale |
| title_fullStr | Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale |
| title_full_unstemmed | Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale |
| title_short | Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale |
| title_sort | malaria diagnosis from pooled blood samples comparative analysis of real time pcr nested pcr and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large scale |
| topic | malaria real-time PCR nested PCR Plasmodium diagnosis |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762011000600008&lng=en&tlng=en |
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