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1996-02-26
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Document 1066
DOCN M9621066
TI Nested polymerase chain reaction for diagnosis and monitoring treatment
response in AIDS patients with tuberculous meningitis.
DT 9602
AU Scarpellini P; Racca S; Cinque P; Delfanti F; Gianotti N; Terreni MR;
Vago L; Lazzarin A; Infectious Diseases Department, San Raffaele
Scientific; Institute, Milan, Italy.
SO AIDS. 1995 Aug;9(8):895-900. Unique Identifier : AIDSLINE MED/96014963
AB OBJECTIVE: To investigate the usefulness of polymerase chain reaction
(PCR) from cerebrospinal fluid (CSF) for rapid diagnosis and assessing
treatment response of tuberculous meningitis (TBM) in AIDS patients.
PATIENTS: Forty-four CSF samples from 10 patients with TBM confirmed by
autopsy or by a culture of CSF (41 samples) and from two patients with
highly probable TBM (three samples) were analysed. CSF specimens were
collected before and during standard antituberculous treatment. CSF
samples from 24 AIDS patients with autopsy evidence of other neurologic
diseases were studied as controls. METHODS: A nested PCR amplifying a
123 base-pair fragment of the IS6110 sequence was developed. Heating to
95 degrees C for 15 min was used for pre-PCR treatment of samples.
RESULTS: Detection limit was 10(2) colony-forming units per ml or 10 fg
purified Mycobacterium tuberculosis DNA. M.tuberculosis DNA was detected
in CSF from all the 12 confirmed or highly probable TBM cases. CSF was
positive by nested PCR in 17 of 17 (100%) and 18 of 27 (67%) samples
collected before and during therapy, respectively. Clinical and
microbiological follow-up > or = 2 weeks was available for seven
patients. PCR-positive CSF converted to M. tuberculosis DNA negative in
four patients that showed improvement during treatment, but it remained
positive in three patients who died of disseminated tuberculosis. All
the CSF samples from the non-TBM controls were negative by nested PCR.
CONCLUSIONS: Nested PCR for detection of M. tuberculosis DNA is specific
for diagnosis of TBM and more sensitive than conventional bacteriology.
Moreover, nested PCR could be a useful method for assessing treatment
response in AIDS patients with TBM.
DE AIDS-Related Opportunistic Infections/*DIAGNOSIS/MICROBIOLOGY Base
Sequence Case Report DNA Primers/GENETICS DNA,
Bacterial/CEREBROSPINAL FLUID/GENETICS Evaluation Studies Human
Molecular Sequence Data Mycobacterium tuberculosis/GENETICS/ISOLATION &
PURIF Polymerase Chain Reaction/*METHODS/STATISTICS & NUMER DATA
Sensitivity and Specificity Tuberculosis,
Meningeal/*COMPLICATIONS/*DIAGNOSIS/MICROBIOLOGY JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).