Rapid detection of Neisseria meningitidis in cerebrospinal fluid by one-step polymerase chain reaction of the nspA gene

A polymerase chain reaction (PCR) protocol for the rapid detection of meningococcal DNA in cerebrospinal fluid (CSF) was developed and optimized. A set of primers based on Neisseria surface protein A ( nspA) gene sequence was designed to amplify a 481-bp product specific for N. meningitidis. We test...

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Bibliographic Details
Published in:	Diagnostic microbiology and infectious disease Vol. 51; no. 2; pp. 85 - 90
Main Authors:	de Filippis, Ivano, do Nascimento, Carlos R.S., Clementino, Maysa B.M., Sereno, Andrea B., Rebelo, Cristina, Souza, Nadjla N.F., Riley, Lee W.
Format:	Journal Article
Language:	English
Published:	New York, NY Elsevier Inc 01-02-2005 Elsevier
Subjects:	Bacteriology Base Sequence Biological and medical sciences Cohort Studies DNA, Bacterial - analysis DNA, Bacterial - cerebrospinal fluid Female Fundamental and applied biological sciences. Psychology Humans Infectious diseases Male Medical sciences Meningitis, Meningococcal - cerebrospinal fluid Meningitis, Meningococcal - diagnosis Meningococcal DNA Microbiology Miscellaneous Molecular Sequence Data Neisseria meningitidis Neisseria meningitidis - genetics Neisseria meningitidis - isolation & purification Neisseriaceae nspA gene Polymerase Chain Reaction Rapid diagnostic Sensitivity and Specificity Time Factors Viral Nonstructural Proteins - analysis Viral Nonstructural Proteins - genetics nspA gene Rapid diagnostic Neisseria meningitidis Meningococcal DNA Microbiology Neisseriaceae Cerebrospinal fluid Infection Polymerase chain reaction Gene Bacteriosis Bacteria Micrococcales Detection Meningococcal disease
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Summary:	A polymerase chain reaction (PCR) protocol for the rapid detection of meningococcal DNA in cerebrospinal fluid (CSF) was developed and optimized. A set of primers based on Neisseria surface protein A ( nspA) gene sequence was designed to amplify a 481-bp product specific for N. meningitidis. We tested 85 N. meningitidis strains obtained from patients with meningococcal meningitis and 112 CSF samples from patients with suspected meningococcal meningitis. No amplification of the nspA gene was observed from other Neisseriaceae species (except from N. gonorrhoeae) and from other bacteria frequently associated with meningitis. N. meningitidis belonging to different serogroups yielded the same product after PCR amplification. The sensitivity and specificity of our protocol was determined by comparing the results of specific amplification of nspA gene by PCR reaction ( nspA-PCR) with those obtained by conventional methods. All positive samples by conventional methods were confirmed by nspA-PCR, whereas 48% of negative samples after culture and latex agglutination tested positive by nspA-PCR. The use of nspA-PCR proved to be a rapid diagnostic method, in which sensitivity and specificity may not be affected by prior antibiotic treatment.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0732-8893 1879-0070
DOI:	10.1016/j.diagmicrobio.2004.10.004