Persistence of dna in cell cultures may jeopardize the analysis of human herpesvirus 6 dynamics by means of realtime pcr. The quantitative range of this assay is 500 to 2,000,000 hhv6 dna. The detection of human herpesvirus 6 hhv 6 dna in peripheral blood mononuclear cells pbmc, throat swabs, and cerebrospinal fluid csf was determined using the polymerase chain reaction pcr. We report a case of unilateral optic disc edema and retinal vasculitis associated with hhv 6 infection. Herpesviridae is a large family of dna viruses that cause infections and certain diseases in animals, including humans. The assay sensitivity was 10 copies of dna per well, with a linear dynamic. Pcr dna test on plasma or serum qualitative or quantitative yes. Chimeric enzymes may be used as surrogate to develop and evaluate new inhibitors. Human herpesvirus 6 hhv 6 latently infects a majority of adults. Structure analysis of dna polymerases may help for rational design of novel drugs. Realtime quantitative pcr for human herpesvirus 6 dna ncbi.
Component test codes cannot be used to order tests. Herpesvirus dna polymerases are attractive targets for development of inhibitors. Qualitative and quantitative polymerase chain reaction pcr for human herpesvirus 6. Ebv, cytomegalovirus cmv, and human herpesvirus 6 hhv6 dna. Herpesvirus 6 hhv 6 dna, quantitative realtime pcr this test is used to determine the presence of hhv 6 dna in patients specimens. Human herpesvirus 6associated uveitis with optic neuritis. Pdf multicenter comparison of pcr assays for detection. Hhv 6 exhibits a wide cell tropism in vivo and, like other herpesviruses, induces a lifelong latent infection in humans. In some cases, additional time should be allowed for additional confirmatory or additional reflex tests. This test looks for human herpesvirus 6 hhv6 by screening for viral dna in the blood. However, the absence of hhv6 dna in the plasmaserum does not mean that there isnt a lowlevel persistent infection in the tissues e. Laboratory and clinical aspects of human herpesvirus 6. Nested polymerase chain reaction pcr was used to analyse csf samples from 365 consecutive hiv infected patients with neurological symptoms.
In all, 104 blood samples were obtained 3 times during pregnancy and once at delivery. Csf, plasma, whole blood qualitative pcr dna test on whole blood is useless for differentiating active from latent hhv 6 7 infection. Jan 18, 2008 quantitative pcr assays with a standard thermal cycling profile permitting simultaneous detection of herpes simplex virus hsv, varicella zoster virus vzv, cytomegalovirus cmv, epstein. Methods seven consecutive patients hospitalized with dress were enrolled in a prospective study to evaluate evidence of active hhv 6 infection. Isolated in 1986 during attempts to find novel viruses in patients with lymphoproliferative diseases, hhv 6 is now recognized as a tcell lymphotropic virus with high affinity for cd4 lymphocytes. The sensitivity of the assay is very dependent upon the quality of the specimen submitted. Realtime pcr for quantification of human herpesvirus 6 dna. Successful after admission, her first cerebrospinal fluid.
A realtime quantitative pcr assay has been developed to measure human herpesvirus 6 hhv 6 dna in biological specimens. Using this assay, we quantitated the amount of hhv6 or hhv7 dna in. Human herpesvirus6associated acute lymphadenitis in. The results obtained should be used in conjunction with clinical findings to make accurate diagnosis. Background there is a current debate regarding the association of human herpesvirus 6 hhv 6 infection and drug reaction with eosinophilia and systemic symptoms dress. These methods rely on the subjective decision of the assessor, obstructing the ability to obtain unanimous results. We have developed and validated an alternative tcid50 readout approach where infection in the titration culture plate is assessed by. The role of human herpesvirus 6 hhv 6 in disease beyond primary infection remains unclear.
The objective was to evaluate the frequency of human herpesvirus 6 hhv 6 dna detection in the csf of patients infected with hiv and its relation to brain disease and systemic hhv 6 infection. Intra and interassay variations were, respectively, 0. Pcr followed by sequencing of dna extracted from the lymph nodes identified the virus as hhv 6, type b. We have developed and validated a new reverse transcription pcr rt pcr assay for hhv 6 that can determine the presence of hhv 6 in clinical specimens and differentiate. Primary human herpesvirus 6 hhv 6 and 7 hhv7 infections were identified in febrile children by qualitative and quantitative polymerase chain reaction pcr assays. Detection of human herpesvirus 6 dna in clinical samples of. Multicenter comparison of pcr assays for detection of human herpesvirus 6 dna in serum. Dna from 711 ad and control brains was analyzed for pcr reactivity to hhv6a andor hhv6b hhv6 demonstrated little specificity to ad brains over controls by either method these complementary methods do not support strong association between hhv6 and ad. Currently clinicians rely mainly on blood lymphocyte culture and serological tests to diagnose hhv. Early diagnosis of primary human herpesvirus 6 infection in. Fhv6d herpesvirus 6 hhv6 dna, quantitative realtime pcr download test. Prolonged therapy with antivirals may lead to emergence of drugresistant isolates. Human herpesvirus 6 hhv 6, which is usually responsible for exanthem subitum in children, can be reactivated from its latent state. Pdf realtime pcr for quantification of human herpesvirus 6.
These closely related viruses are two of the nine herpesviruses known to have humans as their primary host. A positive result indicates the presence of specific dna from human herpesvirus 6 hhv 6 and supports the diagnosis of infection with this virus. Human herpesvirus 6 in cerebrospinal fluid of patients. The polymerase chain reaction pcr was carried out on the plasma or sera of patients to determine the value of pcr in the diagnosis of hhv. The diagnosis of human herpesvirus 6 hhv 6 infection represents a complex issue because the most widely used diagnostic tools, such as immunoglobulin g antibody titer determination and qualitative dna pcr with blood cells, are unable to distinguish between latent clinically silent and active often clinically relevant infection. Human herpesviruses 6 and 7 and central nervous system. Sep 17, 2018 a nested panherpesvirus polymerase chain reaction pcr targeting a region of the dna polymerase gene was used to test 129 western european hedgehogs from across great britain, 20112016. Active human herpesvirus 6 infection in patients with. We found hhv6 dna in the peripheral blood mononuclear cells of 53. Detection of human herpesvirus 6 by reverse transcriptionpcr. Realtime quantitative pcr for human herpesvirus 6 dna. Turnaround time is defined as the usual number of days from the date of pickup of a specimen for testing to when the result is released to the ordering provider. A realtime quantitative pcr assay has been developed to measure human herpesvirus 6 hhv6 dna in biological specimens.
In about 1% of the population hhv 6 exists in a chromosomally integrated form cihhv 6 that resides in every somatic and germ cell and can be transmitted through the germ line. Pdf realtime quantitative pcr for human herpesvirus 6 dna. Detection and characterisation of multiple herpesviruses in. Quantitative pcr for human herpesviruses 6 and 7 journal of. Almost all healthy individuals have low levels of latent hhv 6 in the whole blood. Human herpesvirus 6 hhv 6 is the common collective name for human betaherpesvirus 6a hhv6a and human betaherpesvirus 6b hhv6b. Human herpesvirus 6 detection in alzheimers disease cases. Diagnosis was based on the differential detection of viral dna in peripheral blood mononuclear cells pbmc, but not in saliva. Human herpesvirus6 hhv6 qrtpcr test the test is performed to detect the presence of human herpes virus type 6 hhv6 transcripts in endometrial biopsy samples. Aug 16, 2019 human herpesvirus 6 hhv 6 was the sixth herpesvirus discovered. Human herpesvirus 6 hhv6a and hhv6b by quantitative pcr.
Pdf multicenter comparison of pcr assays for detection of. Epsteinbarr virus, cytomegalovirus, and human herpesvirus 6. Pdf persistence of dna in cell cultures may jeopardize the. Hhv6 is never found in plasma or serum unless there is an acute infection or the individual has cihhv6. Association of human herpesvirus 6 infection with drug. The family name is derived from the greek word herpein to creep, referring to spreading cutaneous lesions, usually involving blisters, seen in flares. The assay sensitivity was 10 copies of dna per well, with a linear dynamic range of 10 to 107 copies of hhv 6 dna. The multiplex assay was used to quantify the ebv, cmv, and hhv6 dna in 46 transplant recipients. Treatment of human herpesvirus 6 encephalomyelitis in immunocompetent patient eric denes, laurent magy, karine pradeau, sophie alain, pierre weinbreck, and sylvie rangerrogez we report a case of human herpesvirus 6 hhv 6 encephalomyelitis in an immunocompetent patient, which was confirmed by viral amplification from cerebrospinal fluid. Quantification of hhv 6 dna is based upon the realtime pcr amplification and detection of hhv 6 genomic dna. Cerebrospinal fluid samples from 245 pediatric patients median age 43 days, submitted for evaluations of possible sepsis or of neurologic symptoms, were tested for hhv 6 and hhv7 dna by polymerase chain reaction.
A healthy 63yearold man noted a decrease in the vision of his left eye. The assay sensitivity was 10 copies of dna per well, with a linear dynamic range of 10 to 10 7 copies of hhv 6 dna. On examination, his left eye had moderate muttonfat keratic precipitates, vitreous. These viruses contain dna surrounded by a lipid envelope. As a noticeable difference with respect to other human herpesviruses, genomic hhv 6 dna is. Reactivation of human herpesvirus 6 hhv 6 and cytomegalovirus cmv during pregnancy and transmission of the viruses to the fetus were investigated by polymerase chain reaction pcr and serology.
Human herpesvirus 6 associated acute lymphadenitis in immunocompetent adults. We have developed a new, highly sensitive, quantitative pcr assay for the accurate measurement of hhv6 dna in tissuederived cell suspensions and body. Labcorp test details for human herpesvirus 6 hhv 6 quantitation, dna pcr. A quantitative pcr assay for the detection of human herpesvirus 6 hhv6 variants. Pcr inhibition and dna extraction efficiency control internal standard validated specimen.
Information on how hhv6 is spread is limited but research has found that the majority of people are infected when they are very young. Reactivation of human herpesvirus 6 during pregnancy the. Rapid quantitative pcr assays for the simultaneous detection. Labcorp test details for human herpesvirus 6 hhv6 quantitation, dna pcr.
A negative result indicates the absence of detectable dna from hhv 6 in the specimen, but it does not negate the presence of the virus or active or recent disease. Pdf persistence of dna in cell cultures may jeopardize. When previously tested and stored dna samples were examined, results of the multiplex realtime pcr assay were as sensitive and speci. Realtime quantitative pcr for human herpesvirus 6 dna article pdf available in journal of clinical microbiology 3811. The role and frequency of human herpesviruses hhv 6 and 7 in central nervous system cns diseases of children are unclear. Multicenter comparison of pcr assays for detection of human. Pcr provides more rapid results than other methods, including culture. In another 107 women, samples were obtained only at delivery. Human herpesvirus 6 hhv 6 is a widespread betaherpesvirus which is genetically related to human cytomegalovirus hcmv and now encompasses two different species. Fhv6d herpesvirus 6 hhv 6 dna, quantitative realtime pcr download test. Herpesvirus6 hhv6 is a member of the herpesviridae family. Dec 18, 2012 for titer assessment of human herpesvirus 6 hhv 6, ifa targeting viral proteins or a tcid50 method with ocular inspection for cpe can be used. Realtime pcr for quantification of human herpesvirus 6. Barr virus ebv, and human herpesvirus 6 hhv6 dna were developed and validated for diagnostic use.
Development and validation of a qpcr based tcid 50 method. Organisms may be detected by pcr prior to detection by immunological methods. Geneproof human herpesvirus 67 hhv67 pcr kit geneproof as. Diagnosis of primary human herpesvirus 6 and 7 infections in.