EQUINE herpesvirus type 1 (EHV-1) infection is widespread in horse populations throughout the world and produces well-documented syndromes of respiratory disease, abortion, neonatal foal death and myeloencephalopathy (van Maanen 2002). The epidemiology of EHV-1 is characterised by lifelong latent infection after primary exposure. The reactivation of a latent stage in a subclinically affected an...

Supplementary material 1 Real-time quantitative PCR: Sequences of the primers and TaqMan probe for ADAMTS1, 4, 5, 8, 9 and 15 were designed using Primer Express software (Applied Biosystems, Foster City, CA): forward primer 5’-CCATCCCAAGAGTATCACATGTCT-3’, reverse primer 5’-CACTATGACACAGCAATTCTTTTCAC-3’ and TaqMan probe 5’-FAM-CCCACACAAGTCCTGTC-MGB-3 for ADAMTS1; forward primer 5’-TCACTGACTTCCTG...

BACKGROUND AND AIM OF THE WORK In recent years, the diagnosis of toxoplasmosis has been improved by Real-time PCR assays. In this study we compared the performances of two Real-time PCRs (FRET and TaqMan protocols) already described in the literature, and one nested-PCR, currently used in our laboratory for the molecular diagnosis of toxoplasmosis. METHODS We evaluated the sensitivity and the...

In this article, the development of a new TaqMan-based one-step real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay for detection and quantification of Crimean-Congo hemorrhagic fever virus (CCHFV) RNA is described. Selected oligos targeting the highly conserved S region of CCHFV were designed by using our oligo design and analysis software, Oligoware 1.0. None of the prime...

Nosocomial infections caused by Acinetobacter spp. resistant to carbapenems are increasingly reported worldwide. Carbapenem-resistant Acinetobacter (CRA) is becoming a serious concern with increasing patient morbidity, mortality, and lengths of hospital stay. Therefore, the rapid detection of CRA is essential for epidemiological surveillance. Polymerase chain reaction (PCR) has been extensively...

BACKGROUND Hepatitis B virus (HBV) DNA levels are crucial for managing chronic hepatitis B (CHB). It was unclear whether Daan real-time polymerase chain reaction test (Daan test) or COBAS TaqMan HBV DNA Test (Cobas TaqMan) was superior in measuring different HBV DNA levels in clinical specimens. METHODS We enrolled 67 treatment-naïve, HBV surface antigen-positive CHB patients (high baseline v...

Heartwater is a tick borne disease that affects ruminants and wild animals in Africa south of the Sahara. It is caused by Ehrlichia ruminantium and transmitted by the tick Amblyomma hebraeum. The protocols currently used to detect heartwater take several days to complete. Here, we describe the development of a pCS20 quantitative real-time PCR TaqMan probe assay to detect E. ruminantium in lives...

Recently we evaluated a custom TaqMan array card (TAC) detection system, formerly known as a TaqMan low-density array (TLDA) card, for simultaneous real-time PCR identification of 21 pathogens and three control targets in duplicate from respiratory specimens (M. Kodani et al., J. Clin. Microbiol. 49:2175-2182, 2011). We engineered an adaptable and expandable system of in vitro RNA transcripts t...

The polymerase chain reaction (PCR), first described in 1985, is a highly sensitive and specific technique used for the detection of nucleic acids [55]. The inventor of this technology earned a Nobel prize for his achievement [43,44], which has revolutionised research and diagnostic possibilities. Qualitative PCR is a well established and straightforward technology, but quantification of specif...

The article describes a new technology for real-time polymerase chain reaction (PCR) detection of nucleic acids. Similar to Taqman, this new method, named Snake, utilizes the 5'-nuclease activity of Thermus aquaticus (Taq) DNA polymerase that cleaves dual-labeled Förster resonance energy transfer (FRET) probes and generates a fluorescent signal during PCR. However, the mechanism of the probe cl...