Bovine viral diarrhoea virus (BVDV) is an important pathogen of dairy cattle. In this study, bulk milk samples representing a total of 4105 milking cows, from 18 dairy cattle herds in the suburb of Mashhad- Iran, were tested for presence of BVDV by the use of a nested reverse transcription polymerase chain reaction (Nested RT- PCR) assay. Non of the cows in the herds had been vaccinated against...

AIMS To evaluate immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR) for melanoma associated antigens (MAA) in detecting occult melanoma cells in lymph nodes which were missed on routine pathology. METHODS Occult melanoma cells were sought in 436 lymph nodes from 32 patients with cutaneous melanoma of the lower extremities by immunohistochemistry using the melano...

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...

We developed a sensitive and rapid real-time reverse transcription-polymerase chain reaction (RT-PCR) assay to detect influenza A H5N1 virus in clinical samples. This assay was evaluated with samples from H5N1-infected patients and demonstrated greater sensitivity and faster turnaround time than nested RT-PCR.

To examine the limitations of reverse transcription (RT)-PCR for the detection of circulating tumor cells in blood, we established a RT-PCR for carcinoembryonic antigen (CEA). Whole blood (10(7) nucleated cells) was mixed with cells from the colon cancer cell line LS174T (concentrations ranging from 0 to 10(6) cells). RT-PCR was performed to detect CEA mRNA in blood under various conditions. He...

A combination of single-tube reverse transcription (RT)-PCR and nested PCR was used to identify spring viraemia of carp rhabdovirus (SVCV) in infected cell cultures and fish tissues. Two pairs of specific primers (external and internal) were selected from the glycoprotein gene sequence. A specific product of 470 bp was amplified from RNA derived from 34 SVCV isolates including the reference str...

BACKGROUND The control of dengue depends solely on the control of the insect vector and efficient diagnosis of human cases as no vaccines or specific treatments are currently available. Existing diagnostic methods for suspected clinical cases are complicated by the short duration of viraemia and by serological cross-reactivity with epitopes from other flaviviruses. OBJECTIVES To evaluate PCR-...

Background Kyasanur Forest Disease (KFD), a tick borne flavivirus, which was earlier endemic to Karnataka state, India, has been confirmed and detected from neighboring states of Tamil Nadu, Maharashtra, Goa and Kerala states in India. Increased human and vector surveillance therefore becomes essential for the identification of KFD affected regions and control of further spread of the disease. ...

RNase Protection Assay (RPAs) is a highly sensitive and reproducible method of quantitating the levels of specific mRNA transcripts. The introduction of the commercially available Multiprobe RPAs allow comparing and quantifying the expression of up to different mRNA species in a single sample of 1-20 micrograms of total RNA. To generate probes which are not commercially available, we prepared h...