Various approaches can now be taken for amplification of RNA transcripts using the polymerase chain reaction (PCR). Here, we compare three such methods: (i) uncoupled reverse transcription (RT)-PCR (using separate reactions for cDNA synthesis and PCR), (ii) continuous RT-PCR (in which RT and DNA amplification occur in an uninterrupted reaction) using either a single enzyme for both RT and DNA a...

The high complexity and cost of polymerase chain reaction-based molecular diagnostics sometimes limits their use in the clinical diagnostics setting. A new helicase-based isothermal amplification method offers an alternative to standard polymerase chain reaction, allowing amplification and detection of specific DNA sequences at a constant reaction temperature without thermocycling equipment. He...

Introduction: The novel coronavirus disease 2019 (COVID-19) has huge impact on public health. RT-PCR of respiratory samples is generally accepted confirmatory test which can miss several cases due to various factors. Case description: A 32-years-old male without any co-morbidity presented with complaints cough and fever was negative for Reverse Transcription Polymerase Chain Reaction (RT-PCR) t...

Canine distemper virus (CDV) is the cause of canine distemper (CD) which is a severe and highly contagious disease in dogs. In the present study, a duplex reverse transcription polymerase chain reaction (RT-PCR) method was developed for the detection and differentiation of wild-type and vaccine strains of CDV. Four primers were designed to detect and discriminate the two viruses by generating 6...

Background and Aims: Influenza virus is a major pathogen involved in respiratory illnesses during winter seasons. A variety of diagnostic methods have been developed to identify influenza viruses in clinical specimen. Methods: Nasal and pharyngeal samples taken from patients were inoculated into Madin-Darby canine kidney (MOCK) cells and embryonated chicken eggs (ECEs). The culture media was a...

A rapid identification of dengue viruses from clinical samples by using a nested reverse transcriptase-polymerase chain reaction (RT-PCR) procedure was carried out for diagnostic and epidemiological purposes. RT-PCR identified DEN-1 and DEN-2 viruses in 41% (41/100) of previously confirmed cases and provided an accurate confirmation of DHF in four fatal cases. RT-PCR was also useful for detecti...

We passaged 52 serum samples from dengue patients on C6/36 cells for 7 days and checked the culture fluids by RT-PCR. Two serum samples, which were negative by direct RT-PCR, became positive. One sample was collected on fever day 1 and the other on fever day 2. Results indicate that combination of reverse transcriptase-polymerase chain reaction (RT-PCR) with passage of serum samples on C6/36 ce...

dear editor-in-chief we read with interest the study by khazaei et al. (1) in which the authors have nicely concluded that pcr is more sensitive test than ziehl-neelsen staining and histo-pathological examination for the diagnosis of tuberculosis (tb). they have rightly pointed to use pcr, selectively, in acidfast bacilli negative paucibacillary forms of tb. however, we intend to highlight few ...

An assay based on reverse transcription coupled with the polymerase chain reaction (RT-PCR) was used for the detection of hog cholera virus (HCV) and bovine virus diarrhoea virus (BVDV) in cell culture. In this study, a precipitate of the supernatants derived from cell cultures infected with HCV and BVDV was used in RT reactions, in place of extracted viral RNA. Both RT and PCR were performed u...