Determination of Viral Load by Quantitative Real-Time PCR in Herpes Simplex Encephalitis Patients

Bhullar, Shradha S.; Chandak, Nitin H.; Purohit, Hemant J.; Taori, Girdhar M.; Daginawala, Hatim F.; Kashyap, Rajpal S.

doi:10.1159/000351521

Cited by 31 publications

(20 citation statements)

References 21 publications

(21 reference statements)

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“…In support of this assumption, (i) we found no correlation between HSV PCR C T values, WBC counts, and protein levels, regardless of whether pooled or PCR assayspecific C T values were considered in the analysis. In line with this, the ranges of CSF WBC counts and protein levels were comparable between patients with confirmed CNS HSV infections displaying high (Ͼ10,000 copies/ml) HSV DNA content in CSF and those with low content (Ͻ10,000 copies/ml), as determined by a quantitative real-time PCR assay (20). (ii) The overall median HSV PCR C T values were not significantly different between CSF specimens from patients that met the Reller criteria and those from patients that did not; nevertheless, the scant number of cases in the latter group limits the conclusions that can be made.…”

Section: Discussionsupporting

confidence: 78%

Missing Cases of Herpes Simplex Virus (HSV) Infection of the Central Nervous System When the Reller Criteria Are Applied for HSV PCR Testing: a Multicenter Study

et al. 2019

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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 2 years of age and are not immunocompromised, the so-called Reller criteria. In this multicenter study, we retrospectively assessed the validity of these screening criteria in our setting. A total of 4,404 CSF specimens submitted for HSV PCR testing to the respective microbiology laboratories at the participating hospitals between 2012 and 2018 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 seven were deemed to truly have HSV encephalitis. Overall, no significant correlation between HSV PCR cycle threshold (C T ) values and WBC counts or total protein levels was found. In addition, median HSV PCR C T s were comparable between patients who met the Reller criteria and those who did not (P ϭ 0.531). In summary, we show that HSV DNA may be detected in CSF specimens with normal WBC and protein levels collected from immunocompetent individuals older than 2 years with HSV encephalitis. Nevertheless, the data also indicate that the number of cases detected could be lowered at least by half if CSF specimens with borderline WBC counts (4 cells/mm 3 ) as well as children of any age are systematically tested. KEYWORDS central nervous system infections, cerebrospinal fluid, herpes simplex virusH erpes simplex virus (HSV) is among the commonest etiological agents of sporadic central nervous system (CNS) infection in immunocompetent patients (1). Timely diagnosis and prompt initiation of acyclovir are crucial for patient survival and recovery (1). Requests for cerebrospinal fluid (CSF) HSV PCR testing, the diagnostic gold standard (1), can be a burden on molecular microbiology laboratory resources; moreover, the results are often noncontributory. Based on previous studies (2, 3), Hanson and Cerebrospinal fluid specimens. 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 in this study. No restrictive acceptance criteria for testing were in place at any of the centers. Specifically, 1,474 specimens were received at

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Section: Discussionsupporting

confidence: 78%

Missing Cases of Herpes Simplex Virus (HSV) Infection of the Central Nervous System When the Reller Criteria Are Applied for HSV PCR Testing: a Multicenter Study

et al. 2019

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“…According to a Swedish report, 29 cases of acute HSV encephalitis were assayed by quantitative FQ-PCR. After analyzing the relationship between disease course and viral load, they found that the higher level of viral load in CSF, the graver disease severity and the longer disease course [10]. However, our study revealed no relevance between viral load and hospitalization duration.…”

Section: Discussioncontrasting

confidence: 74%

“…It is reported in the literature that the magnitude of viral load was closely correlated with disease severity [9]. And early diagnosis and early treatment could lower the viral load and boost the therapeutic efficacies [10]. In recent years, the arrival of fluorescence quantitative-polymerase chain reaction (FQ-PCR) has marked a great transformation from qualitative to quantitative analyses of viruses.…”

Section: Introductionmentioning

confidence: 99%

Clinical Epidemiological and Pathogenic Analyses for Acute Viral Infections of Central Nervous System in Hospitalized Children of Hunan Province, China

Zhang¹,

Tian²,

Zhu³

et al. 2018

Clin Microbiol

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Background: Virus infection of the central nervous system is a common disease in children. The major causes of ineffective therapy are misdiagnosis and delayed interventions. Based upon our pilot study, consecutive specimens of cerebrospinal fluid were collected and detected. Methods: From October 2012 to July 2014, a total of 161 cerebrospinal fluids samples were collected from the patients with a clinical diagnosis of viral encephalitis at our hospital. The technique of real-time fluorescence quantitative-polymerase chain reaction was employed.Results: Viruses were detected in 71 cases with an overall detection rate of 44.10%. In our cohort, the average viral load was 198.24 ± 993.61 copies/μL. And the viral loads had no obvious correlations with hospitalization duration. Enterovirus had the highest detection rate of 18.01% and occurred predominantly in spring and summer. The highest detection rate was found in the 3-6 year age group. Most cases of Human herpesviruses-6 were detected in autumn. And Adenovirus positive cases were largely restricted for rural children. Only 1 Mumps virus positive case had an enlargement of parotid gland. None of 12 Measles virus positive cases had the classic manifestations of measles. Conclusions:During the study period, 11 common viruses in cerebrospinal fluids were detected from hospitalized children of viral encephalitis. The detection rates have slightly increased as compared with earlier reports. And Measles virus and Mumps virus were increasingly detected. For positive and negative viral cases, no differences existed in gender, age, urban-rural region or clinical characteristics. And the positive cases of Enterovirus, Measles virus and Mumps virus have their unique profiles.

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“…Total DNA was extracted from 200 µl of CSF samples from patients by using a ZR viral DNA kit (Zymo Research, USA), according to the manufacturerʹs protocol. The amplification reactions were carried out using SYBR green and the primers for HSV which were described earlier (Bhullar et al, 2013). The cycle threshold (Ct) was calculated as the cycle number at which the …”

Section: Methodsmentioning

confidence: 99%

Identification of an immunodominant epitope in glycoproteins B and G of herpes simplex viruses (HSVs) using synthetic peptides as antigens in assay of antibodies to HSV in herpes simplex encephalitis patients

Ss¹,

Nh²,

Nn³

et al. 2014

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Summary. -Herpes simplex encephalitis (HSE) is a severe viral infection of the central nervous system (CNS). Assay of antibody response is widely used in diagnostics of HSE. The aim of this study was to identify an immunodominant epitope determining the antibody response to herpes simplex viruses (HSVs) in cerebrospinal fluid (CSF) of HSE patients. The synthetic peptides that resembled type-common as well as type-specific domains of glycoproteins B (gB) and G (gG) of these viruses were evaluated for binding with IgM and IgG antibodies in CSF samples from HSE and non-HSE patients in ELISA. The QLHDLRF peptide, derived from gB of HSV was found to be an immunodominant epitope in the IgM and IgG antibody response. The patients with confirmed and suspected HSE showed in ELISA against this peptide 26% and 23% positivities for IgM, 43% and 37% positivities for IgG and 17% and 15% for both IgM and IgG antibodies, respectively. The total positivities of 86% and 75% for both IgM and IgG antibodies were obtained in the patients with confirmed and suspected HSE, respectively. These results demonstrate that a synthetic peptide-based diagnostics of HSE can be an efficient and easily accessible alternative. This is the first report describing the use of synthetic peptides derived from HSVs in diagnostics of HSE using patientsʹ CSF samples.

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Determination of Viral Load by Quantitative Real-Time PCR in Herpes Simplex Encephalitis Patients

Cited by 31 publications

References 21 publications

Missing Cases of Herpes Simplex Virus (HSV) Infection of the Central Nervous System When the Reller Criteria Are Applied for HSV PCR Testing: a Multicenter Study

Missing Cases of Herpes Simplex Virus (HSV) Infection of the Central Nervous System When the Reller Criteria Are Applied for HSV PCR Testing: a Multicenter Study

Clinical Epidemiological and Pathogenic Analyses for Acute Viral Infections of Central Nervous System in Hospitalized Children of Hunan Province, China

Identification of an immunodominant epitope in glycoproteins B and G of herpes simplex viruses (HSVs) using synthetic peptides as antigens in assay of antibodies to HSV in herpes simplex encephalitis patients

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