Ocorrência de resultados falso-negativos na reação de PCR em Tempo Real (PCR-TR) no diagnóstico da doença meningocócica / False-negative results in Real-Time PCR (RT-PCR) formeningococcal disease diagnosis
Rev. Inst. Adolfo Lutz (Online)
;
72(1): 65-71, 2013. tab
Article
in English
| LILACS, SES-SP
| ID: lil-729390
ABSTRACT
Factors responsible for false-negative results (F-N) in RT-PCR assay for detecting N. meningitidis in serumand CSF samples were investigated. As the meningococcal disease should be rapidly treated because ofits high mortality and epidemic potential, the F-N in diagnostic testing may cause treatment failuresand/or on disease restraint in community. Thus, it is crucial to learn the factors which cause F-N in RTPCRassays. The F-N were induced by inhibition, low quantity of target DNA in extracted samples, andinadequate temperature employed at PCR annealing procedure. As bacterial DNA concentration in samplesmight be highly variable, the ideal sample volume to be extracted could not be defined. As previouslyrecommended for N. meningitidis gene-grouping by RT-PCR assay, the annealing temperature at 60 °Cwas not suitable for B and W135 genogroups. Altogether, these factors induced F-N in 31 samples; therefore,30 % of N. meningitidis detected by RT-PCR were classified as non-genogrouped. The inhibitors and/orthe low amount of target DNA induced F-N on RT-PCR, independently of the specimen volume used forextracting DNA. However, adjustments on the PCR annealing temperature and amount of extracted DNAadded into the reaction might avoid the occurrence of the majority of F-N.
Full text:
Available
Index:
LILACS (Americas)
Main subject:
Cerebrospinal Fluid
/
Diagnostic Techniques and Procedures
/
False Negative Reactions
/
Real-Time Polymerase Chain Reaction
/
Neisseria meningitidis
Type of study:
Diagnostic study
/
Screening study
Language:
English
Journal:
Rev. Inst. Adolfo Lutz (Online)
Year:
2013
Type:
Article
Institution/Affiliation country:
Instituto Adolfo Lutz/BR
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