Respiratory viruses are among the most important causes of human morbidity and mortality worldwide, especially for infants and young children. In the past years, a few commercial multiplex RT-PCR assays have been used to detect respiratory viruses in spite of the high cost. In the present study, an improved single-tube multiplex reverse transcription PCR assay for simultaneous detection of 13 r...

BACKGROUND Staphylococcus aureus is often responsible for fatal infections and recent upsurge of resistant strains has resulted in therapeutic failure. The identification of this microorganism is a major challenge to medical microbiologists in developing countries. METHODS One hundred and eighty five isolates which had been previously isolated from the nares of 185 healthy college students' v...

Polydimethyl siloxane (PDMS)–glass microchip has a very strong surface effect on polymerase chain reaction (PCR), leading to a very poor PCR yield. In the work reported here, practical dynamic passivation of surfaces of PDMS–glass microchip using polyethylene glycol (PEG) or polyvinylpyrrolidone (PVP) was achieved using a conventional thermocycler. The passivation procedure was cost-effective a...

evidences from many studies suggested a polymerase chain reaction (pcr) as a valuable method for diagnosing infectious disease in the transplant recipients. we used this method for detection of toxoplasma, gondii and human cytomegalovirus in blood specimens from patients after bone marrow or kidney transplantation. dna of both infectious agents were detected using two separate sets of nested pr...

A novel method, which involves a nested PCR in a single closed tube, was developed for the sensitive detection of Erwinia amylovora in plant material. The external and internal primer pairs used had different annealing temperatures and directed the amplification of a specific DNA fragment from plasmid pEA29. The procedure involved two consecutive PCRs, the first of which was performed at a high...

One-tube nested PCR was developed for diagnosis of pulmonary tuberculosis using sequences based on thel6SrRNA gene. The usage of primers 16SOL, 16SOR, 16SIL and 16SIR with optimized conditions could detect 555 bp DNA band from 21 species, 41 strains of mycobacteria and one isolate of Nocardia asteroides. It also revealed a specific 306 bp DNA band from 59 strains of M. tuberculosis complex. Cro...

Diagnosis of Rift Valley fever (RVF) is based on serology and virus isolation. The disadvantages of the former include poor sensitivity, high cost, risks associated with using infectious virus as antigen, the lengthy duration of ELISA as well as cross-reactivity with other Phleboviruses. We developed, optimised and evaluated a one-tube reverse-transcription-polymerase chain reaction (RT-PCR) fo...

The use of PCR amplification to detect target DNA sequences has many applications in plant genotyping, gene mapping, diagnostics, and diversity assessment. PCR itself is simple to set up, and requires little hands-on time. However, most methods for preparing DNA from plant tissues are time consuming, tedious, and labor intensive. Virtually all require mechanical disruption, such as grinding in ...

A rapid, inexpensive, simple, and accurate multiplex polymerase chain reaction (PCR) was developed in a single tube for identification of Mycobacterium tuberculosis. Assessment of sensitivity and specificity of simple PCR was performed with 116 strains of M. tuberculosis complex (MTC) and 144 strains of nontuberculous mycobacteria (NTM) compared with the biochemical method. Specific amplificati...