Herpes Simplex Virus Genotyping in Neurological Abnormalities-Clinical Relevance for Disease Monitoring (original) (raw)
Related papers
The Brazilian Journal of Infectious Diseases, 2011
Diagnosis of herpes simplex encephalitis (HSE) is based on the detection of herpes simplex virus (HSV) DNA in patients' CSF samples. HSV DNA quantitation has the potential for estimating the effects of antiviral therapy. The aim of this study was to diagnose HSV DNA in HSE suspected patients and the quantitative analysis of its genome using real-time PCR to assess the value of the viral load in the course of antiviral treatment. The CSF samples were collected from 236 consecutive HSE suspected patients from November 2004 to May 2008. Upon DNA extraction, the samples were analyzed by Real-Time PCR assay. A set of primers amplified a common sequence of HSV glycoprotein B gene. The copy numbers of unknown samples were expressed via a standard curve drawn with a known amount of amplified cloned plasmid. Of the 236 samples, 137 (58%) came from males and 99 (42%) from females. The HSV genome was detected in 22 (9.3%) patients by PCR, 13 males/ 9 females. Serial CSF samples were available from 10 of the 22 patients. The range of the HSV DNA copy numbers in the clinical samples ranged from 2.5 × 10 2 to 1.7 × 10 6 copies/mL of CSF. Quantitative PCR results can be helpful in evaluating the efficacy of antiviral therapy in the above-mentioned patients. There is an association between the initial viral load and the duration of treatment course.
VirusDisease, 2019
Herpes simplex viruses (HSVs) cause a latent infection in humans which is mainly associated with characteristic cold sores or fever blisters and genital blisters. Large segments of the world population are suffering from the HSV infection and early diagnosis as well as treatments are needed to avoid further complications. HSV surveillance is very sparse, especially from developing countries including India. The aim of the present study is to develop and evaluate molecular assays for rapid detection and typing of HSV. In the present study, viral DNA was extracted from cerebro-spinal fluid from HSV suspected encephalitis patients. The conventional multiplex PCR for HSV-1 and HSV-2 was optimized and their comparative analysis was done with Real-Time qPCR for detection and typing of HSV. Out of 137 clinical samples, eleven samples (8.03%) were diagnosed as HSV positive by Real-Time qPCR while ten (7.3%) by conventional multiplex PCR which were further typed as subtyping HSV-1 (nine) and HSV-2 (two). Real-Time qPCR is highly sensitive and able to detect 9.4 9 10 1 to 3.1 9 10 6 copies/ml of HSV DNA. Conventional PCR was found to be having 99.21% specificity with 100% sensitivity. The positive predictive value was 90.91% whereas negative predictive value was 100%. Logistic regression indicates blisters with pain and skin rash as the most significant symptoms associated with HSV infection. The present study could be applied for rapid, specific, sensitive and cost-effective diagnosis of HSV-1 and HSV-2 thereby helpful in better patient management through early detection and treatment of HSV.
Establishment of PCR for the early diagnosis of herpes simplex encephalitis
Journal of Medical Virology, 1990
The early detection of herpes simplex encephalitis (HSE) represents a major problem in viral diagnostics. We have now established a test system based on PCR for the specific amplification of HSV-DNA in cerebrospinal fluid (CSF) samples followed by oligonucleotide hybridization. The test proved to be very sensitive. Five molecules of HSV 1 DNA still yielded positive signals after hybridization. The assay was applied to CSF samples from 6 patients with confirmed HSE. All except one CSF sample obtained from the 2nd to the 8th day after onset of neurological symptoms yielded positive results. The primers used also exhibit a certain degree of crossreactivity with HSV 2, as revealed by testing of a CSF sample from an HSV2-infected child. No positive signals were obtained with human DNA and with DNA from CMV-and VZV-infected fibroblasts. Also 42 CSF samples of patients suffering from other diseases of the CNS yielded negative results. The present results indicate that the detection of HSV DNA in CSF by PCR represents a valuable tool for the early diagnosis of HSE.
PCR in herpes simplex virus infections of the central nervous system
Serodiagnosis and Immunotherapy in Infectious Disease, 1994
Herpes simplex virus (HSV) infections of the central nervous system (CNS) together comprise a special group of herpetic diseases with respect to residual damage and difficulties in diagnosis. The various syndromes and manifestations are presented, in particular the most common and severe form, herpes simplex virus encephalitis (HSE). The probable reasons for the failure of routine viral methods for establishing a rapid and early diagnosis in HSE are discussed. The technical problems of polymerase chain reaction (PCR) in the diagnosis of HSE and its use as a diagnostic method are reviewed.
Detection of Herpes simplex virus DNA by real-time PCR
Journal of clinical microbiology, 2000
Molecular detection of herpes simplex virus (HSV) DNA is recognized as the reference standard assay method for the sensitive and specific diagnosis of central nervous system infections caused by HSV. In this study, a molecular assay based on real-time PCR on the LightCycler (LC) instrument was evaluated and compared with a home-brew molecular assay. The detection limit of the LC assay was determined with 10-fold dilutions of plasmid pS4 with the SalI restriction fragment of the DNA polymerase gene and with the First European Union Concerted Action HSV Proficiency Panel. A total of 59 cerebrospinal fluid (CSF) specimens were investigated for the comparative study. With plasmid pS4, the detection limit of the LC assay was found to be 10(4) copies per ml, i.e., 12.5 copies per run. When samples of the First European Union Concerted Action HSV Proficiency Panel were tested, 2x10(3) to 5x10(3) HSV type 1 genome equivalents (GE) per ml, i.e., 2.5 to 6.3 GE per run, could consistently be d...
Journal of clinical …, 2001
A sensitive multiplex PCR assay for single-tube amplification that detects simultaneous herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), varicella-zoster virus (VZV), human cytomegalovirus (CMV), and Epstein-Barr virus (EBV) is reported with particular emphasis on how the method was optimized and carried out and its sensitivity was compared to previously described assays. The assay has been used on a limited number of clinical samples and must be thoroughly evaluated in the clinical context. A total of 86 cerebrospinal fluid (CSF) specimens from patients which had the clinical symptoms of encephalitis, meningitis or meningoencephalitis were included in this study. The sensitivity of the multiplex PCR was determined to be 0.01 and 0.03 50% tissue culture infective doses/the reciprocal of the highest dilution positive by PCR for HSV-1 and HSV-2 respectively, whereas for VZV, CMV and EBV, 14, 18, and 160 ag of genomic DNA were detected corresponding to 48, 66, and 840 genome copies respectively. Overall, 9 (10.3%) of the CSF samples tested were positive in the multiplex PCR. HSV-1 was detected in three patients (3.5%) with encephalitis, VZV was detected in four patients (4.6%) with meningitis, HSV-2 was detected in one neonate (1.16%), and CMV was also detected in one neonate (1.16%). None of the samples tested was positive for the EBV genome. None of the nine positive CSF samples presented herpesvirus coinfection in the central nervous system. Failure of DNA extraction or failure to remove any inhibitors of DNA amplification from CSF samples was avoided by the inclusion in the present multiplex PCR assay of ␣-tubulin primers. The present multiplex PCR assay detects simultaneously five different herpesviruses and sample suitability for PCR in a single amplification round of 40 cycles with an excellent sensitivity and can, therefore, provide an early, rapid, reliable noninvasive diagnostic tool allowing the application of antiviral therapy on the basis of a specific viral diagnosis. The results of this preliminary study should prompt a more exhaustive analysis of the clinical value of the present multiplex PCR assay.
Detection of Herpes Simplex Virus Infection in Cerebrospinal Fluid
Medical Laboratory Journal, 2016
Background and Objective: Herpes simplex encephalitis is a life-threatening consequence of the central nervous system (CNS) infection with Herpes simplex virus (HSV). Although it is a rare disease, mortality rates reach 70% in the absence of therapy and only a minority of individuals can return to normal function. The aim of this study was to determine possible correlation between HSV infection and the incidence of encephalitis in patients with neurological signs. Methods: Overall, 152 CSF samples were tested from patients with neurological signs referred to Mahdieh Clinical Laboratory in Isfahan from 2010 to 2013. After cerebrospinal fluid (CSF) collection, DNA was extracted and real-time polymerase chain reaction (PCR) was performed for HSV detection. Results: Of 152 patients tested, 50 were diagnosed with encephalitis. HSV DNA was present in the CSF of 13 patients with encephalitis. HSV was significantly higher (p< 0.05) in patients with encephalitis, which shows the significance of infection as an etiological factor of this disease. About 60% of the encephalitis cases were in age range of 1-24 months. Conclusion: According to the findings of the present study, Cesarean section is recommended for HSV-positive mothers. A routine real-time PCR test is suggested for HSV detection in patients with encephalitis to avoid unnecessary antiviral treatments.
Journal of Clinical Microbiology
Previous studies suggested that Herpes simplex virus (HSV) PCR testing can be safely deferred in patients with normal cerebrospinal fluid (CSF) white blood cell (WBC) counts and protein levels as long as they are older than two years of age and are not immunocompromised, the so-called Reller criteria. In this multicenter study, we retrospectively assessed the validity of this screening criteria in our setting. A total of 4,404 CSF specimens submitted to the respective microbiology laboratories at the participating hospitals between 2012 and 2018 for HSV PCR testing were included. Six commercially-available HSV PCR assays were used across the participating centers. Ninety-one of the 4,404 CSF specimens (2.1%) tested were positive for HSV DNA (75 samples for HSV-1 and 16 for HSV-2). Nine patients failed to meet the Reller criteria, of whom 7 were deemed to truly have HSV encephalitis. Overall, no significant correlation between HSV PCR cycle threshold (Ct) values and WBC counts or tot...
Journal of Neurology, Neurosurgery & Psychiatry, 1993
A polymerase chain reaction (PCR) was used to detect herpes simplex virus (HSV) deoxyribonucleic acid (DNA) in CSF of 109 patients with possible herpes simplex encephalitis. HSV DNA was found in 20/109 patients. In 14 of these patients the diagnosis was confirmed by a rise in CSF antibodies, isolation of HSV from the brain, or both. In 3 patients CSF antibodies did not rise and 3 patients did not have a follow up lumbar puncture or a brain biopsy. In 19/20 patients HSV DNA was present in the first CSF specimen. The virus was identified as HSV I in 15 patients and HSV II in 4; the virus was not typed in the other patient. A possible diagnosis of herpes simplex encephalitis was not confirmed in the 89 PCR-negative patients. HSV DNA was present in CSF of 3 patients who had meningitis with herpetic genital infections but it was not found in 24 patients with other neurological diseases. The results suggest that the detection of HSV DNA in CSF using a PCR assay will be an accurate method of early diagnosis of herpes simplex encephalitis.