Introduction:EBV:Epstein-Barr Virus (EBV) is a member of the herpesvirus family (HHV-4), and one of the most common human viruses. In the United States, as many as 95% of adults between 35 and 40 years of age have been infected. Epstein-Barr can cause Infectious mononucleosis, also known as 'glandular fever', 'Mono' and 'Pfeiffer's disease'. There is a body of evidence linking EBV and cancer formation especially in Burkitt's lymphoma and Nasopharyngeal carcinoma.
CMV:Cytomegalovirus (CMV) is the most common congenital infection in the United States. Cytomegalovirus, or CMV is found universally throughout all geographic locations and socioeconomic groups and infects between 50% and 85% of adults in the United States by 40 years of age. CMV infection is more widespread in developing countries and in areas of lower socioeconomic conditions. For most healthy persons who acquire CMV after birth there are few symptoms. However, CMV infection is important to certain high-risk groups. Major areas of concern are; the risk of infection to the unborn baby during pregnancy, the risk of infection to people who work with children, the risk of infection to the immunocompromized person, such as organ transplant recipients and persons infected with human immunodeficiency virus (HIV) and the risk of infection to newborn babies.
BKV:Polyomavirus is the sole genus of viruses within the family Polyomaviridae. Polyomaviruses are DNA-based (double-stranded DNA, ~5000 base pairs, circular genome), small (40-50 nanometers in diameter), icosahedral in shape and do not have a lipoprotein envelope. They are potentially oncogenic; they often persist as latent infections in a host without causing disease but may produce tumors in a host of a different species, or a host with an ineffective immune system. BK virus produces a mild respiratory infection and can affect the kidneys of immunosuppressed transplant patients. In Jan 2008, a new species, Merkel cell polyoma virus, was described as the likely causative agent of Merkel skin cancer. All the polyomaviruses are highly common childhood and young adult infections. Most of these infections appear to cause little or no symptoms. These viruses are probably lifelong persistent among almost all adults. Diseases caused by human polyomavirus infections are most common among persons who become immunosuppressed by AIDS, old age or after transplantation.
Intended Use:The genesig Real-Time PCR Screening Multiplex Kit for Epstein–Barr Virus, Cytomegalovirus and Polyomavirus BK is designed for the in vitro detection of Cytomegalovirus, Epstein-Barr virus and Polyomavirus BK. genesigPLEX qPCR Kits are unique multiplex qPCR assays that allow simultaneous real-time PCR detection of different preselected viruses. Individual primers and probes have been designed permitting multiple virus detection in a single reaction through separate fluorescent channels. Human genesigPLEX qPCR Kits also all include a human endogenous control to confirm extraction of a valid biological template.
Specificity:The genesigPLEX kit is designed for the in vitro detection of Epstein Barr Virus (EBV also known as HHV4), Cytomegalovirus (CMV also known as HHV5) and Polyomavirus BK (BKV). The primers and probe sequences in this kit have 100% homology with a broad range of clinically relevant reference sequences based on a comprehensive bioinformatics analysis. They therefore have a very broad detection profile.
Dynamic Range of Test:Under optimal PCR conditions genesig kits have very high priming efficiencies of >90% and can detect between 1x10e8 and 1x10e2 copies of target template.
Suitable Sample Material:All kinds of sample material suited for PCR amplification can be used. Please ensure the samples are suitable in terms of purity, concentration, and RNA/DNA integrity. Always run at least one negative control with the samples. To prepare a negative control, replace the template DNA sample with RNase/DNase free water.
Template preparation buffer, for resuspension of positive control template
RNase/DNase free water, for resuspension of primer/probe mix
Reagents and Equipment to be Supplied by the User:Real-Time PCR InstrumentDNA extraction kitThis kit is recommended for use with genesig Easy DNA/RNA extraction kit
However, it is designed to work well with all processes that yield high quality DNA with minimal PCR inhibitors
Pipettors and TipsVortex and CentrifugeThin walled 1.5ml tubesqPCR plates
Storage and Stability:This kit is stable at room temperature but should be stored at -20ºC on arrival. Once the lyophilized components have been resuspended they should not be exposed to temperatures above -20ºC for longer than 30 minutes at a time and unnecessary repeated freeze/thawing should be avoided. The kit is stable for six months from the date of resuspension under these circumstances.
