Molecular techniques based on the detection of genomic sequences by reverse transcription (RT)-PCR, nested PCR, or real-time PCR have made possible the rapid diagnosis of dengue virus (DENV) infections, and these approaches have been accepted by clinical laboratories as the new standard method for the detection of dengue virus in acute-phase serum samples. One of these PCR-based assays, the two...

AIMS To develop a highly sensitive and rapid protocol for simultaneous detection and differentiation of Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) in pepper and tomato. In this study, we use the multiplex PCR technique to detect dual infection of these two viruses. METHODS AND RESULTS A multiplex RT-PCR method consisting of one-tube reaction with two primer pairs targeted to re...

A dried blood spot (DBS) is a well-accepted means for the collection, transport, and storage of blood samples for various epidemiologic, serologic, and molecular assays for human immunodeficiency virus (HIV) studies. It is particularly important for mother-to-infant-transmission studies of affected individuals living in remote areas. We have developed a real-time PCR method to detect HIV type 1...

We have developed a single-tube nested real-time PCR (STN-RT PCR) assay using the repetitive, transposon-like element IS1111 as the DNA target to facilitate early diagnosis of acute Q fever. The use of our proposed diagnostic procedures, including IgM detection by serology and the STN-RT PCR assay, significantly increased the diagnostic sensitivity for Q fever to 78%, compared to 29% when serol...

One hundred and ninety eight samples of human and animal sources from different areas in El-Sharkia Governorate were used in this study to isolate Staphylococcus species and to determine the presence of coagulase ( coa) gene,a virulence gene detected in S. aureus (CoPS), by PCR. Primary isolation on mannitol salt agar, $-haemolysis, tube coagulase test (TCT) and other biochemical characterizati...

BACKGROUND A significant role of Renin-angiotensin system (RAS) genetic variants in the pathogenesis of essential hypertension and cardiovascular diseases has been proved. This study aimed to develop a new, fast and cheap method for the simultaneous detection of two missense single nucleotide polymorphisms (T207M or rs4762 and M268T orrs699) of angiotensinogen (AGT) in single-step Multiplex Hex...

PCR has been used as an aid in the diagnosis of invasive aspergillosis for almost 2 decades. A lack of standardization has limited both its acceptance as a diagnostic tool and multicenter clinical evaluations, preventing its inclusion in disease-defining criteria. In 2006, the European Aspergillus PCR Initiative was formed. The aim of the initiative was to provide optimal standardized protocols...

Drug-resistant Mycobacterium tuberculosis can be rapidly diagnosed through nucleic acid amplification techniques by analyzing the variations in the associated gene sequences. In the present study, a locked nucleic acid (LNA) probe-based real-time PCR assay was developed to identify the mutations in the rpoB gene associated with rifampin (RFP) resistance in M. tuberculosis. Six LNA probes with t...

We report the first description of osteomyelitis due to Bartonella henselae genotype I in an immunocompetent middle-aged woman. The diagnosis was established by serology, histopathology, and PCR analysis of osseous and lymph node tissues. The mycobacteria growth indicator tube inoculated with the lymph node aspirate was used for PCR analysis.

Sequencing in less than four lanes on a gel, using only one fluorescent label for the four bases (without variations in electrophoretic mobilities or problems with spectral overlap) in one tube was introduced in (1). In that process T7 DNA polymerase and buffer containing magnesium ions were used. By adjusting the ddNTPs ratios the sequence is deduced from the peak magnitudes corresponding to t...