2013
DOI: 10.1590/s0365-05962013000100025
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The use of nested Polymerase Chain Reaction (nested PCR) for the early diagnosis of Histoplasma capsulatum infection in serum and whole blood of HIV-positive patients*

Abstract: The aim of the study was to detect the rDNA sequences and their regions in Histoplasma capsulatum, which could be considered species-specific and used as a molecular method for this diagnosis by the technique of nested polymerase chain reaction (nested PCR), employing specific sequences (primers) for H. capsulatum: 18S rDNA region (HC18), 100 kDa (HC100) and the sequence 5.8 S-ITS rDNA (HC5.8). The PCR sequences HC18, HC100 and HC5.8 resulted in a specificity of 100%. The molecular assays may increase the spec… Show more

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Cited by 24 publications
(22 citation statements)
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“…Similar to the antigen testing, PCR presented excellent analytical performance, displaying a similar accuracy in comparison to the antigenuria-based assays [30]. By reviewing studies that compared primers, it was possible to see that Hcp100 is the most widely targeted gene and has shown better methodological performance [23,24].…”
Section: Discussionmentioning
confidence: 96%
See 1 more Smart Citation
“…Similar to the antigen testing, PCR presented excellent analytical performance, displaying a similar accuracy in comparison to the antigenuria-based assays [30]. By reviewing studies that compared primers, it was possible to see that Hcp100 is the most widely targeted gene and has shown better methodological performance [23,24].…”
Section: Discussionmentioning
confidence: 96%
“…Most studies in the literature evaluated the performance of nested PCR in the diagnosis of DH, a method that is associated with the increased risk of amplicons contamination, since it involves the manipulation of amplified products. Many of the PCR tests for histoplasmosis were developed in-house [11][12][13][16][17][18][19][20][21][22][23][24][25][26][27]. The gene encoding a 100 kDa molecular protein, which is described as necessary for the survival of H. capsulatum in human cells, has been the most common target for the diagnosis of histoplasmosis [11,12,16,19,23,24,27].…”
Section: Overview Of the Performance Of Molecular Methods In The Diagmentioning
confidence: 99%
“…In recent years, numerous molecular methods have been developed and evaluated to improve the identification of endemic mycoses, including real-time PCR assays for blastomycosis and histoplasmosis [3032], multiplex PCR for histoplasmosis [33], nested PCR assays for identification of histoplasmosis in HIV-infected patients [34], multiplex suspension array identification of multiple species [35], and PCR identification of sporotrichosis [36]. Parallel efforts to map the genomes of important fungal pathogens continue to inform the rational development of these diagnostic methods.…”
Section: New Diagnostic Toolsmentioning
confidence: 99%
“…для исследования в пцр клинического материала от больных гистоплазмозом предложено несколько молекулярных мишеней [28][29][30]. чаще используется уникальный фрагмент гена, кодирующего белок в 110 кда (нср100).…”
unclassified
“…чаще используется уникальный фрагмент гена, кодирующего белок в 110 кда (нср100). этот белок, по-видимому, включается в процессы регуляции, необходимые для адаптации и выживаемости гриба внутри клеток макрофагов [29]. при исследовании клинических образцов чувствительность метода составила 100 %, а специфичность -95,2 % [31].…”
unclassified