Quantitative real-time PCR has become the method of choice for measuring mRNA transcription. Recently, fast PCR protocols have been developed as a means to increase assay throughput. Yet it is unclear whether more rapid cycling conditions preserve the original assay performance characteristics. We compared 16 primer sets directed against Epstein-Barr virus (EBV) mRNAs using universal and fast P...

In view of the importance of early diagnosis of tuberculous meningitis (TBM), the efficiency of the polymerase chain reaction (PCR), one of the most reliable and sensitive DNA-based assays, was compared with conventional methods (acid-fast microscopy and culture) for the detection of M. tuberculosis in cerebrospinal fluid (CSF) specimens from patients suspected of TBM. Of the 29 CSF specim...

Paratuberculosis (John’s disease) is infectious and chronically progressive granulomatous disease which affects domestic and wild ruminants. The causative agent is Mycobacterium avium paratuberculosis (MAP), a slow growing mycobactin dependent acid-fast bacillus. We investigated the detection and frequency of MAP in apparently healthy dromedary and Bactrian camels by insertion...

In diagnostic and research bacteriology settings with budget and staff restrictions, fast and cost-effective genome extraction methods are desirable. If not inactivated properly, cellular and/or environmental DNA nucleases will degrade genomic material during the extraction stage, and therefore might give rise to incorrect results in PCR experiments. When crude cell extracts, proteinase K–treat...

in diagnostic and research bacteriology settings with budget and staff restrictions, fast and cost-effective genome extraction methods are desirable. if not inactivated properly, cellular and/or environmental dna nucleases will degrade genomic material during the extraction stage, and therefore might give rise to incorrect results in pcr experiments. when crude cell extracts, proteinase k–treat...

The Centers for Disease Control and Prevention and United States Army Research Institute for Infectious Diseases have developed real-time PCR assays for the detection of bioterrorism threat agents. These assays all rely on a limited number of approved real-time PCR master mixes. Because the availability of these reagents is a critical element of bioterrorism preparedness, we undertook a joint n...

in view of the importance of early diagnosis of tuberculous meningitis (tbm), the efficiency of the polymerase chain reaction (pcr), one of the most reliable and sensitive dna-based assays, was compared with conventional methods (acid-fast microscopy and culture) for the detection of m. tuberculosis in cerebrospinal fluid (csf) specimens from patients suspected of tbm. of the 29 csf specimens f...

conclusions concurrent pulmonary nocardiosis and tuberculosis is frequent in hiv-infected patients. rapid and sensitive methods such as pcr are recommended for detection of such co-infections. results from 157 sputum specimens, positive samples by acid fast staining, culture and pcr for m. tuberculosis were reported for 7.6% (12/157), 10.1% (16/157) and 7% (11/157) of samples, respectively. no ...

A novel subtype of influenza A pandemic virus, A(H1N1)v, was recently reported by the Centers for Disease Control and Prevention (CDC) (Atlanta, GA) and WHO in April 2009 (1). Infection most commonly results in a mild respiratory tract infection, but there are reports of severe cases with mortalities even in patients without chronic diseases (2). Early detection of infection is important in pro...

Current study evaluated the hsp65 Nested PCR Restriction Fragment Length Polymorphism Analysis (hsp65 Nested PCR-PRA) to detect and identify Mycobacterium tuberculosis complex directly in clinical samples for a rapid and specific diagnosis of tuberculosis (TB). hsp65 Nested PCR-PRA was applied directly to 218 clinical samples obtained from 127 patients suspected of TB or another mycobacterial i...