Real-time nested multiplex PCR for the detection of herpes simplex virus types 1 and 2

The goal was to show that a multiplex, high-throughput genome-sequencing approach is feasible for simultaneously sequencing seven HSV-1 ocular strains. Eighty-one were from genitourinary medicine (GUM) patients and the other 68 specimens were from other patients with skin lesions. The mechanistic basis for this repression is unknown, but its global nature suggests regulation by an epigenetic mechanism such as DNA methylation. Melting peaks of HSV-1, HSV-2, and VZV positive samples by melting curve analysis. Methods: Forty specimens, collected from 33 patients with clinically suspected herpes virus ocular infection, were tested. Forty specimens, collected from 33 patients with clinically suspected herpes virus ocular infection, were tested. The goal was to show that a multiplex, high-throughput genome-sequencing approach is feasible for simultaneously sequencing seven HSV-1 ocular strains.

Its performance was compared to those of PCR assays routinely used in the laboratory, namely, a laboratory-developed test for HSV DNA on the LightCycler instrument and a test using a commercial master mix for VZV DNA on the ABI7500fast system. HSV-1/2 multiplex assay: Luminex bead sets were covalently linked with specific HSV-1 and -2 recombinant antigens (gG1 and gG2), purified whole HSV-1 and -2 antigens, and a goat-anti-human IgG serving as an internal control. The sensitivity of detection of each of the viruses using the LightCycler assay was compared to that of the conventional assay using external quality assessment material. A multiplex real-time PCR (quantitative PCR [qPCR]) assay detecting herpes simplex virus (HSV) and varicella-zoster virus (VZV) DNA together with an internal control was developed on the BD Max platform combining automated DNA extraction and an open amplification procedure. We performed multiplex polymerase chain reaction (PCR) assay to examine multiple viral reactivations simultaneously in 24 patients undergoing auto-HSCT between September 2010 and December 2012. The FilmArray® (BioFire Diagnostics, LLC, Salt Lake City, Utah) is a user-friendly, fully automated, multiplex PCR system that is being developed for rapid point-of-care use. A sensitive multiplex polymerase chain reaction was achieved by optimization of parameters such as the primers, magnesium, and dNTPs concentrations.


Multiplex Real-Time PCR for detection of pathogen genes by TaqMan® technology. A multiplex real-time PCR test detecting HSV1, HSV2 and Treponema pallidum was designed, validated and evaluated for a period of 6 months on patients attending the Sandyford Initiative (a series of genitourinary medicine clinics in and around Glasgow). A multiplex PCR assay using novel primer sets for cytomegalovirus (CMV), herpes simplex virus (HSV), varicella zoster virus (VZV), and Toxoplasma gondii was developed. HSV-1 or VZV DNA was detected in 13 specimens by simple PCRs (HSV-1 or VZV PCR) and in 12 specimens by multiplex PCR. The sensitivity of detection of each of the viruses using the LightCycler assay was compared to that of the conventional assay using external quality assessment material. Your library or institution may give you access to the complete full text for this document in ProQuest. Your library or institution may give you access to the complete full text for this document in ProQuest.

It is therefore necessary to have a rapid diagnostic test that differentiates between the different causes of infectious conjunctivitis. Methods A multiplex PCR assay using novel primer sets for cytomegalovirus (CMV), herpes simplex virus (HSV), varicella zoster virus (VZV), and Toxoplasma gondii was developed. Adenovirus infection is associated with nosocomial outbreaks and HSV may result in episodes of recurrent ocular inflammation. Multiplex Real-Time PCR for detection of pathogen genes by TaqMan® technology. Your library or institution may give you access to the complete full text for this document in ProQuest. This study aimed to develop a multiplex PCR method for the simultaneous screening of clinical samples for the presence of the 10 viruses presently considered as the major viral causes of acute encephalitis/ encephalopathy in Asia. The infectious agent has only one serotype, but it was classified by restriction enzyme analysis of the whole genome into four genotypes, named I to IV.

The assay, termed MeningoFinder, showed an accordance of 97%, concordance of 96%, interlaboratory sensitivity of 90%, and interlaboratory specificity of 94% compared to PCRs. Results were compared to a consensus standard, defined as the result obtained by at least 2 of the 3 molecular methods. To compare this system against the bioMérieux NucliSens easyMAG using a range of common clinical sample types. To test this hypothesis, we devised a PCR-based strategy to simultaneously screen for adenovirus 12 (Ad12), cytomegalovirus (CMV), and herpes simplex viruses (HSV) 1 and 2 in formalin-fixed paraffin-embedded small bowel biopsies from adult and childhood CD patients. Multiple studies have shown that GUDs are strongly associated with the transmission and the acquisition of HIV infection. An internal amplification control was included in the mPCR reaction. Herpes simplex virus 1 (HSV-1) is a well-adapted human pathogen that can invade the peripheral nervous system and persist there as a lifelong latent infection.

Gone are the days of expensive, laborious, and subjective testing for laboratories that process large numbers of samples.

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