herpes
Laboratory diagnosis of central nervous system infections caused by herpes viruses.

Sauerbrei A, Wutzler P.

Institute for Antiviral Chemotherapy, Friedrich-Schiller University Jena, Winzerlaer Stasse 10, D-07745 Jena, Germany. andreas.sauerbrei med.uni-jena.de

BACKGROUND: Herpesviruses may be associated with various types of central nervous system (CNS) infections. Herpes simplex encephalitis (HSE) has to be considered one of the most severe diseases. As effective antiviral drugs are available, rapid and reliable diagnosis has become important. OBJECTIVES: To describe polymerase chain reaction (PCR) and serological methods for the detection of herpes virus-induced CNS infections by the example of HSE. STUDY DESIGN: 620 cerebrospinal fluid (CSF) and 2400 serum samples from 2700 selected hospitalized patients with clinical suspicion of encephalitis were tested for herpes simplex virus (HSV) as well as varicella-zoster virus (VZV) DNA and HSV-specific antibodies, respectively. RESULTS: HSV-1 DNA could be detected in eight and HSV-2 in three patients with focal encephalitis. In addition, HSV-2 DNA was found in two newborns with encephalitis and two adults suffered from transverse lumbar myelitis. One VZV DNA-positive patient had developed herpes zoster accompanied by meningoencephalitis, and in the other an encephalitis without cutaneous rash was diagnosed. Intrathecal antibody synthesis could be measured when CSF was cleared from viral DNA. CONCLUSIONS: The detection of viral DNA by PCR technique has become the "gold standard" method for laboratory diagnosis of herpes virus infections of CNS. Serodiagnosis may be useful to confirm the diagnosis retrospectively.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12091081&dopt=Abstract herpes medicine



herpes
The genome of a very virulent Marek's disease virus.

Tulman ER, Afonso CL, Lu Z, Zsak L, Rock DL, Kutish GF.

Plum Island Animal Disease Center, Agricultural Research Service, U. S. Department of Agriculture, Greenport, New York 11944, USA.

Here we present the first complete genomic sequence, with analysis, of a very virulent strain of Marek's disease virus serotype 1 (MDV1), Md5. The genome is 177,874 bp and is predicted to encode 103 proteins. MDV1 is colinear with the prototypic alphaherpes virus herpes simplex virus type 1 (HSV-1) within the unique long (UL) region, and it is most similar at the amino acid level to MDV2, herpes virus of turkeys (HVT), and nonavian herpes viruses equine herpes viruses 1 and 4. MDV1 encodes 55 HSV-1 UL homologues together with 6 additional UL proteins that are absent in nonavian herpes viruses. The unique short (US) region is colinear with and has greater than 99% nucleotide identity to that of MDV1 strain GA; however, an extra nucleotide sequence at the Md5 US/short terminal repeat boundary results in a shorter US region and the presence of a second gene (encoding MDV097) similar to the SORF2 gene. MD5, like HVT, encodes an ICP4 homologue that contains a 900-amino-acid amino-terminal extension not found in other herpes viruses. Putative virulence and host range gene products include the oncoprotein MEQ, oncogenicity-associated phosphoproteins pp38 and pp24, a lipase homologue, a CxC chemokine, and unique proteins of unknown function MDV087 and MDV097 (SORF2 homologues) and MDV093 (SORF4). Consistent with its virulent phenotype, Md5 contains only two copies of the 132-bp repeat which has previously been associated with viral attenuation and loss of oncogenicity.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10933706&dopt=Abstract herpes medicine



herpes
Detection of herpes virus infection of the CNS: the experience of hospital Geral de Santo Antonio.

Cabeda JM, Castro AP, Amorim ML, Mendes AC, Monteiro L, Amorim JM.

Unidade de Biologia Molecular, Hospital Geral de Santo Antonio, R.D. Manuel II, 4099-001 Porto, Portugal. jcabeda mail.telepac.pt

BACKGROUND: PCR detection of CSF Herpes virus DNA is an important tool in the diagnosis of CNS infections. Use of this test has been shown to have an impact on patient management as measured by shortened patient stays, specific therapeutic intervention, reduction of empirical expensive therapy administration, all of which should translate into significant health care savings. OBJECTIVE: The present study aimed at implementing, and evaluating both clinically and analytically the performance of several commercially available PCR based assays for the detection of Herpes virus infections of the CNS. STUDY DESIGN: A total of 314 patients with suspected CNS Herpesvirus infection were investigated, between 1999 and 2001, by Stair primers PCR. Starting on January 2002, two commercially available real-time-PCR systems were implemented and tested using the Stair primers PCR assay as golden standard and three external control proficiency panels along with serial dilutions of positive clinical samples. RESULTS: Sensitivity of the assay was determined to be <200 copies per ml for HSV and <1250 copies per ml for CMV. Positive results were obtained for 17 patients (6 HSV-1, 1 HSV-2, 1 EBV, 1 CMV, 6 VZV and 2 HHV-6) whose clinical and analytical findings were consistent with the PCR results. A real-time-PCR procedure was introduced in 2002 with similar sensitivity, but a more rapid response. CONCLUSION: Conventional end-point PCR proved useful to the diagnosis of CNS herpes virus infection, with an impact on the clinical intervention. However, the use of Real-Time-PCR has greatly enhanced these advantages, making results available at a much earlier time, thus significantly reducing the need for empirical treatment.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12091082&dopt=Abstract herpes medicine



herpes
Detection of herpes virus genomes by polymerase chain reaction in cerebrospinal fluid and clinical findings.

Minjolle S, Arvieux C, Gautier AL, Jusselin I, Thomas R, Michelet C, Colimon R.

Laboratoire de Bacteriologie-Virologie, Universite Rennes 1, 2 avenue, du Pr Leon Bernard, CS 34317, 35 043 Rennes cedex, France.

BACKGROUND: The viruses of the Herpesviridae family, in particular herpes simplex virus types 1 (HSV-1) and 2 (HSV-2), cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella zoster virus (VZV), and human herpes virus 6 (HHV-6), are responsible for numerous infections of the central nervous system (CNS). These infections manifest as diverse clinical signs, many of which are not specific. The diagnosis of these infections is necessary to make it possible to adapt treatment appropriately, as treatment is specific for the particular virus concerned. OBJECTIVES: To apply a polymerase chain reaction (PCR) technique for the diagnosis in a single reaction of the six herpes viruses most frequently detected in the cerebrospinal fluid (CSF) and to analyse clinical events in patients presenting positive results in PCR for herpes viruses. STUDY DESIGN: We studied 141 patients, from whom 180 CSF samples were collected. The clinical files of the patients were consulted retrospectively, and a list of clinical signs was recorded. After testing by targeted PCR, at the clinician's demand, we tested these samples by herpes consensus PCR, which detects six herpes viruses (HSV-1, HSV-2, CMV, EBV, VZV, HHV-6), in a single PCR. RESULTS: Targeted PCR tests identified 25 CSF samples (13.9%), corresponding to 18 patients (12%), as positive. The herpes consensus PCR test detected 49 samples (27.2%) as positive, resulting in the identification of 54 individual viruses (four samples displayed co-infection) from 39 patients (27%). 130 CSF samples, from 101 patients, tested negative by both techniques. 23 HIV-positive patients (30.6%), three HIV-negative immunocompromised patients (27%), and 14 immunocompetent patients (25%) were CSF PCR-positive. In HIV-positive patients, CMV was the virus most frequently identified (13%), followed by EBV (10.6%), VZV (5.3%) and finally HSV-1 and HSV-2 (both 1.3%). We did not detect HHV-6 in any of these samples. We detected only HSV-2, EBV and VZV in the 11 HIV-negative immunocompromised patients. CSF samples of immunocompetent patients contained mostly VZV (9%) and HSV-1 (7.3%). CONCLUSIONS: The herpes consensus PCR for a given virus was more sensitive than the standard, targeted PCR used in our laboratory. The clinical signs presented by patients infected with HSV-1, HSV-2 and CMV were similar to those reported in previous studies. For VZV, we report the possibility of mild, transient cerebral viral reactivation. Our data on the detection of EBV by PCR suggest that the PCR test is of predictive value for cerebral lymphoma in immunocompromised patients. The possible role of HHV-6 in a subacute neurological disorder merits further investigation.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12091083&dopt=Abstract herpes medicine



herpes
An appraisal of screening for maternal type-specific herpes simplex virus antibodies to prevent neonatal herpes.

Rouse DJ, Stringer JS.

Center for Research on Women's Health, Department of Obstetrics and Gynecology, University of Alabama at Birmingham, USA.

Almost all neonatal herpes simplex virus infections occur as a result of first-episode maternal infection during late pregnancy when delivery occurs before the development of protective maternal antibodies. Screening of pregnant women for the presence of type-specific herpes simplex virus antibodies has therefore been suggested as a means of identifying women vulnerable to herpes simplex virus acquisition and subsequent transmission of herpes simplex virus to their neonates. Couples in whom herpes simplex virus serotype discordance is identified could be counseled regarding sexual behavior modification to avoid maternal herpes simplex virus infection. However, the ramifications of routine screening for herpes simplex virus susceptibility during pregnancy could be profound in terms of costs, prenatal care delivery, and even social duress. The recent US Food and Drug Administration approval of type-specific herpes simplex virus antibody assays for clinical use lends temporal urgency to the need for a critical examination of the relevant data. After we performed such an evaluation and created a decision analysis model to assess the potential cost-effectiveness of herpes simplex virus antibody screening, we concluded that screening for maternal type-specific herpes simplex virus antibodies cannot be recommended to prevent neonatal herpes.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10942477&dopt=Abstract herpes medicine



herpes
Herpes Consensus PCR test: a useful diagnostic approach to the screening of viral diseases of the central nervous system.

Calvario A, Bozzi A, Scarasciulli M, Ventola C, Seccia R, Stomati D, Brancasi B.

Virology Laboratory, Hygiene, Epidemiology and Public Health Department, Policlinico, P.za G. Cesare, 11-70124 Bari, Italy. m.quarto igiene.uniba.it

BACKGROUND: Infections of the central nervous system (CNS) are a difficult diagnostic problem for both clinicians and microbiologists. Various clinical signs, such as encephalitis, myelitis, meningitis, may be associated with herpes viruses. The use of multiplex 'Herpes Consensus' polymerase chain reaction (HC-PCR) in association with nested PCR (nPCR), in addition to classical techniques, made it possible to optimise the management of cerebrospinal fluid (CSF) and serum samples from patients affected by these viral diseases of the CNS. OBJECTIVES: To test by HC-PCR by nPCR and cell culture the CSF and sera from patients with viral infections of the CNS. STUDY DESIGN: We analysed 320 CFS, 154 serum samples and 11 various samples from 286 patients with clinically suspected encephalitis, meningitis or other diseases of the CNS by HC-PCR, nPCR and traditional investigations (cell culture and serological tests). RESULTS: On molecular analysis with the HC-PCR test, 51 CFS samples (15.9%) were positive for at least one of the six target Herpes viruses: fourteen for Herpes simplex 1 (HSV-1), seven for HSV-2, 12 for Cytomegalovirus (CMV; one of which was from an HIV-positive patient), five for Epstein-Barr virus (EBV; four of which were from HIV-positive patients), three for Varicella-Zoster virus (VZV), five for Human Herpes virus type 6 (HHV-6), three for HSV-1 with HHV-6 co-infection (two cases) and HSV-2 co-infection (one case), and two for HHV-6 with CMV or EBV co-infection (both from patients with immune deficiency). A further 12 samples were positive in nPCR for HHV-7 (8), ADV (1), Enterovirus (1), HSV-1 (1), EBV (1). Of the 154 serum samples, 17 (11.0%) tested positive by HC-PCR for HSV-1 (4), HSV-2 (1), CMV(1), EBV(1), VZV(3) or HHV-6(6), 1 with co-HSV-2/VZV infection. A further five samples tested positive for HHV-7 in nPCR. Culture and tests for antibodies did not supply sufficiently sensitive and specific data. CONCLUSIONS: Our laboratory experience shows that herpes viruses play a central aetiological role in viral infections of the CNS. PCR analysis, especially the HC-PCR test, have revolutionised the diagnostic approach to such infections, making possible rapid, specific and highly sensitive baseline screening. In this way, microbiological investigations can lead to prompt diagnosis, which was limited in the past to a very small number of cases.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12091084&dopt=Abstract herpes medicine



herpes
Diagnosis of herpes virus infections of the central nervous system.

Aberle SW, Puchhammer-Stockl E.

Institute of Virology, University of Vienna, Austria, Kinderspitalgasse 15, A-1095 Vienna, Austria.

BACKGROUND: Human herpes viruses may cause infections of the central nervous system (CNS). The early diagnosis of herpes virus-associated neurological diseases is of high importance. OBJECTIVES: The objective of this paper is to summarize the experience gained with the diagnosis of herpes virus infections of the CNS at our institute by polymerase chain reaction (PCR)-based assays within the past few years. STUDY DESIGN: A retrospective analysis of herpes virus desoxy ribonucleic acid (DNA)-positive cerebrospinal fluid (CSF) samples was performed, with particular emphasis on data obtained by quantification of virus DNA in CSF by newly established real-time quantitative PCR assays. RESULTS: Herpesviruses were found in 26.6% of all virus-positive CSF samples detected at our institute between 1995 and 2001. The overall broad testing for different herpes viruses from CSF has led to an increase in the detection rate, especially in relation to varicella zoster virus (VZV)-associated CNS disease. The herpes virus DNA load in CSF was investigated by TaqMan real-time PCR assays that were established for the individual herpes viruses. The amount of virus varied among the individual diseases, associated with herpes simplex virus type 1, herpes simplex virus type 2, VZV and cytomegalovirus, while for Epstein-Barr virus and human herpes virus type 6 only low levels of virus were detectable in CSF. CONCLUSIONS: A generally broad testing for different herpes viruses in CSF samples is highly recommended. In addition, determination of the virus DNA level in CSF by quantitative assays seems to be of high importance for elucidating aspects concerning the prognosis of disease, the prediction of distinct CNS manifestations, and possibly the differentiation between specific virus-associated disease and unspecific presence of virus in CSF, especially in immunocompromised patients.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12091085&dopt=Abstract herpes medicine



herpes
Apolipoprotein E (APOE) phenotype and APOE concentrations in multiple sclerosis and acute herpes zoster.

Pirttila T, Haanpaa M, Mehta PD, Lehtimaki T.

Institute for Basic Research in Developmental Disabilities, Staten Island, NY, USA. tuula.pirttila kuh.fi

OBJECTIVES: There are three major isoforms of apolipoprotein E (apoE), namely apoE2, apoE3, and apoE4, that are products of three alleles (epsilon2,epsilon3,epsilon4) at a single gene locus on chromosome 19. It is well known that the presence of apoE4 increases the risk for the development of Alzheimer's disease and atherosclerosis. The aim of the study was to examine if apoE polymorphism or apoE levels contribute to the severity of the disease in patients with multiple sclerosis or the outcome of nerve damage in patients with herpes zoster infection. MATERIAL AND METHODS: We examined apoE phenotype of 105 MS patients and 41 patients with herpes zoster. We also measured serum and cerebrospinal fluid (CSF) levels of apoE from 93 patients with definite MS using enzyme linked immunosorbent assay. RESULTS: There were no differences in apoE allele frequencies in MS or herpes zoster patients compared to the allele frequencies of controls. The levels of serum or CSF apoE did not differ from those of age-matched controls, nor did they correlate with the disease activity. CONCLUSION: We conclude that apoE does not contribute to the activity of MS or the outcome of herpes zoster.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10949525&dopt=Abstract herpes medicine