A long-term assessment of quantitative hepatitis C virus (HCV) testing was performed at the University of Pittsburgh Medical Center. The Quantiplex HCV RNA 2.0 branched-chain DNA (bDNA) assay (Bayer Diagnostics) for hepatitis C viral load determination was used to test 3,471 specimens. bDNA-negative samples were also tested by an in-house qualitative reverse transcriptase (RT)-PCR assay with a ...

Laboratory diagnosis of viral respiratory infections is generally performed by virus isolation in cell culture and immunofluorescent assays. Reverse transcriptase PCR is now recognized as a sensitive and specific alternative for detection of respiratory RNA viruses. A rapid real-time multiplex PCR assay was developed for the detection of influenza A and influenza B viruses, human respiratory sy...

The objective of this study was to compare agar gel immunodiffusion test (AGID) and immunohistochemistry (IHC) with reverse transcriptase-polymerase chain reaction (RT-PCR) in terms of sensitivity and specificity for the detection of infectious bursal disease virus (IBDV). Thirty-five bursal samples collected from filed outbreak of IBD were evaluated by all 3 diagnostic tests. Sensitivity and s...

Severe acute respiratory syndrome (SARS) has caused a major epidemic worldwide. A novel coronavirus is deemed to be the causative agent. Early diagnosis can be made with reverse transcriptase-polymerase chain reaction (RT-PCR) of nasopharyngeal aspirate samples. We compared symptoms of 156 SARS-positive and 62 SARS-negative patients in Hong Kong; SARS was confirmed by RT-PCR. The RT-PCR-positiv...

Viral metagenomic libraries are a promising but previously untapped source of new reagent enzymes. Deep sequencing and functional screening of viral metagenomic DNA from a near-boiling thermal pool identified clones expressing thermostable DNA polymerase (Pol) activity. Among these, 3173 Pol demonstrated both high thermostability and innate reverse transcriptase (RT) activity. We describe the b...

A detection system based on a multiplex reverse transcription (RT) polymerase chain reaction (PCR) was developed to simultaneously identify multiple viruses in the lily plant. The most common viruses infecting lily plants are the cucumber mosaic virus (CMV), lily mottle virus (LMoV), lily symptomless virus (LSV). Leaf samples were collected at lily-cultivation facilities located in the Kangwon ...

The resolving power of RT-PCR is limited by the efficiency of RNA-to-cDNA conversion. Methods to determine this efficiency, using a real-time PCR assay for quantifying AML1-MTG 8 [t(8;21)] fusion gene transcripts, are described. The efficiency is shown to be directly proportional to RNA template levels. The Moloney murine leukemia virus (MMLV) reverse transcriptase enzyme's conversion efficienc...

Laboratory diagnosis of viral respiratory infections is generally performed by virus isolation in cell culture and immunofluorescent assays. Reverse transcriptase PCR is now recognized as a sensitive and specific alternative for detection of respiratory RNA viruses. A rapid real-time multiplex PCR assay was developed for the detection of influenza A and influenza B viruses, human respiratory sy...

Background Noroviruses are a group of viruses that cause gastroenteritis (illness that usually includes diarrhea and/or vomiting) in people. Gastroenteritis means inflammation of the stomach and small and large intestines. Many different viruses can cause gastroenteritis, including rotaviruses; noroviruses; adenoviruses, types 40 and 41; sapoviruses; and astroviruses. Current techniques used fo...

We have attempted a fine characterization of the angiogenic response induced by multiple myeloma endothelial cells (MMEC) by using the chick embryo chorioallantoic membrane (CAM) assay and by reverse transcriptase-polymerase chain reaction (RT-PCR). Results showed that in the CAM assay MMEC induced an angiogenic response comparable to that of a well-known angiogenic cytokine, namely fibroblast ...