DIRECT DETECTION OF ENTEROVIRUS GENOME IN CELL-CULTURE NEGATIVE CEREBROSPINAL FLUID FROM ASEPTIC MENINGITIS CASES IN BRAZIL

Enteroviruses (EV) are among the most common causes of aseptic meningitis. Standard diagnostic techniques are often too slow and lack sensitivity to be of clinical relevance. EV RNA can be detected within 5 h by a commercially available reverse transcription-PCR (RT-PCR) test kit. Cerebrospinal fluid (CSF) samples from 68 patients presenting with aseptic meningitis during a summer outbreak in Switzerland were examined in parallel with cell culture and commercial RT-PCR. RT-PCR was positive in all 16 CSF specimens positive by cell culture (100%). In addition, 42 of 52 (80%) CSF samples negative by cell culture were PCR positive. In 26 of these 42 (62%) patients, viral culture from other sites (throat swab or stool) was also positive. The CSF virus culture took 3 to 7 days to become positive. Echovirus 30 was the type most often isolated in this outbreak. The sensitivity of CSF RT-PCR based on clinical diagnosis during this aseptic meningitis outbreak in patients with negative bacterial culture results was 85%, i.e., considerably higher than the sensitivity of CSF virus culture (24%). We conclude that this commercial RT-PCR assay allows a positive diagnosis with minimal delay and may thus influence clinical decisions.

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Evaluation of a Commercially Available Reverse Transcription-PCR Assay for Diagnosis of Enteroviral Infection in Archival and Prospectively Collected Cerebrospinal Fluid Specimens

Journal of Clinical Microbiology ◽

10.1128/jcm.36.6.1741-1745.1998 ◽

1998 ◽

Vol 36 (6) ◽

pp. 1741-1745 ◽

Cited By ~ 27

Author(s):

Francisco Pozo ◽

Inmaculada Casas ◽

Antonio Tenorio ◽

Gloria Trallero ◽

Jose M. Echevarria

Keyword(s):

Cell Culture ◽

Cerebrospinal Fluid ◽

Aseptic Meningitis ◽

Reverse Transcription ◽

Diagnostic Systems ◽

Rt Pcr ◽

Enteroviral Infection ◽

Reverse Transcription Pcr ◽

Sporadic Cases ◽

Pcr Method

A commercially available reverse transcription (RT)-PCR method (AMPLICOR EV; Roche Diagnostic Systems, Inc., Branchburg, N.J.) was evaluated for detection of enteroviruses in cerebrospinal fluid from patients with neurological disease. This assay was compared with virus isolation in cell culture and an in-house RT-PCR method designed with a nonoverlapping region of the enteroviral genome. A panel of 200 cerebrospinal fluid specimens prospectively collected from patients with a wide variety of neurological symptoms, including 50 patients involved in three different outbreaks of acute aseptic meningitis, was assayed. A second panel of 97 archived cerebrospinal fluid specimens, stored for 2 to 5 years, from patients with aseptic meningitis associated with several enterovirus outbreaks was also studied. From the first panel, enteroviruses were detected in 13 of 50 specimens by cell culture (26%), in 43 of 50 specimens by AMPLICOR EV (86%), and in 46 of 50 specimens by the in-house assay (92%) from patients with aseptic meningitis associated with outbreak and 1 of 29, 3 of 29, and 4 of 29 specimens, respectively, from sporadic cases of aseptic meningitis. The remaining 121 cerebrospinal fluid specimens from patients with other neurological syndromes were negative by all tests. From the second panel, enteroviral RNA was detected by the AMPLICOR test (31 of 97 specimens, 32%) and the in-house assay (39 of 97 specimens, 40%). According to our results, patients with aseptic meningitis should be analyzed for enteroviral infection in cerebrospinal fluid by RT-PCR methods, and the AMPLICOR EV test is a suitable tool for performing such studies. Archival cerebrospinal fluid specimens are less suitable for evaluation of the performance of RT-PCR methods designed for enterovirus detection.

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Isolation of Parainfluenza Virus Type 3 from Cerebrospinal Fluid Associated with Aseptic Meningitis

American Journal of Clinical Pathology ◽

10.1093/ajcp/99.6.705 ◽

1993 ◽

Vol 99 (6) ◽

pp. 705-707 ◽

Cited By ~ 9

Author(s):

Randall D. Craver ◽

Robert S. Gohd ◽

Daniel R. Sundin ◽

John C. Hierholzer

Keyword(s):

Cerebrospinal Fluid ◽

Aseptic Meningitis ◽

Virus Type ◽

Parainfluenza Virus ◽

Type 3

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AN EPIDEMIC OF ASEPTIC MENINGITIS SYNDROME DUE TO ECHO VIRUS TYPE 6

PEDIATRICS ◽

10.1542/peds.29.3.418 ◽

1962 ◽

Vol 29 (3) ◽

pp. 418-431

Author(s):

David T. Karzon ◽

Norman S. Hayner ◽

Warren Winkelstein ◽

Almen L. Barron

Keyword(s):

Cerebrospinal Fluid ◽

Aseptic Meningitis ◽

Virus Type ◽

Alimentary Tract ◽

Febrile Illness ◽

Acute Onset ◽

Etiologic Agent ◽

Abdominal Muscles ◽

Older Children ◽

Echo Virus

The clinical features of 130 cases of aseptic meningitis syndrome associated with ECHO virus type 6 infection were studied. Characteristically, the onset was acute with the development of fever, headache, muscle pains, and vomiting. A biphasic course was present in 8% of the cases. The physical findings included mild to moderate stiffness or spasm of the neck, back, and posterior thigh muscles. Disturbance in sensorium, cranial nerve involvement, and bulbar signs were conspicuously absent. Absence of deep tendon and superficial reflexes occurred in 16% of the cases. Evidence of mild muscle weakness occurred in 39% of cases, more commonly in the axial rather than peripheral muscles. Most of these patients had bilateral weakness of the anterior neck muscles, but included in the group were five with bilateral weakness of the back or abdominal muscles and six with some involvement of the extremities. Neuromuscular changes were mild and frequently questionable. Where follow-up was available, the changes were usually found to be transient. The effect of age upon the clinical picture was analyzed. Only 3 of 130 patients were less than 4 years of age. Young children had a more acute onset and were admitted to the hospital more promptly than older children and adults. The incidence of pleurodynia was 38% in adults and only 5% in patients less than 20 years of age. The cerebrospinal fluid revealed pleocytosis, with lymphocyte counts ranging from a few cells to 930/mm3. The protein values exceeded 40 mg/100 ml in 48% of the cases, the highest value being 102 mg/100 ml. An undifferentiated febrile illness in household associates of patients with aseptic meningitis, characterized by headache, vomiting and muscle pain, was associated with a high recovery rate of ECHO 6 virus. This is presumably the forme fruste of the fully developed aseptic meningitis syndrome. ECHO 6 virus was recovered from four patients in whom there was evidence of neurologic involvement beyond that seen in the aseptic meningitis syndrome. ECHO 6 virus was the probable etiologic agent in two of these cases. A 15-year-old girl with mild encephalitis marked by disorientation, hallucinations, and weakness of the legs yielded ECHO 6 from her alimentary tract. ECHO 6 was recovered from the cerebrospinal fluid and alimentary tract of a 15-year-old boy with features of the Guillain-Barré syndrome.

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Multiplex PCR for direct identification of Campylobacter spp. in human and chicken stools

Journal of Medical Microbiology ◽

10.1099/jmm.0.47220-0 ◽

2007 ◽

Vol 56 (10) ◽

pp. 1350-1355 ◽

Cited By ~ 45

Author(s):

Aisha Al Amri ◽

Abiola C. Senok ◽

Abdulrahman Yusuf Ismaeel ◽

Ali E. Al-Mahmeed ◽

Giuseppe A. Botta

Keyword(s):

Multiplex Pcr ◽

Direct Detection ◽

False Negative ◽

Enteric Bacteria ◽

Enzymic Activity ◽

Campylobacter Coli ◽

Biochemical Tests ◽

Direct Evaluation ◽

Stool Specimens ◽

Culture Negative

Differentiation between Campylobacter jejuni and Campylobacter coli is problematic in clinical specimens due to fastidious growth requirements and limited biochemical tests. This study describes a rapid, multiplex PCR protocol for the direct detection and differentiation of C. jejuni and C. coli in stools. An evaluation was carried out of this multiplex protocol based on the detection of cadF (genus specific), and hipO (C. jejuni) and asp (C. coli) genes, using stool from patients with Campylobacter enteritis and chicken. Protocol sensitivity was assessed and specificity determined using a panel of enteric bacteria, and evaluation of 30 diarrhoeic stool specimens culture negative for Campylobacter. Of the 114 specimens (54 human and 60 chicken) evaluated by the protocol, 70 (61.4 %) were identified as C. jejuni, 35 (30.7 %) as C. coli and 9 (7.9 %) as a mixed infection/colonization with both species. All mixed infections were identified as C. jejuni by culture. Among the stool specimens that were culture negative for Campylobacter, two (6.7 %) were C. jejuni positive by multiplex PCR. The protocol sensitivity limit was 0.015–0.016 ng C. jejuni and C. coli DNA μl−1 in the specimen. There was no cross-reaction with the reference strains assessed. Comparison of hippurate test and multiplex PCR demonstrated 17 isolates with false-positive hippurate enzymic activity and 7 with false-negative activity. This rapid protocol (turnaround time 6 h) is highly sensitive and specific for direct evaluation of stool for these pathogens. It has significant application for routine clinical diagnostic and epidemiological purposes.

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Increased Levels of Cytokines in Cerebrospinal Fluid of Children with Aseptic Meningitis Caused by Mumps Virus and Echovirus 30

Scandinavian Journal of Immunology ◽

10.1111/sji.12131 ◽

2013 ◽

Vol 79 (1) ◽

pp. 68-72 ◽

Cited By ~ 15

Author(s):

A. Sulik ◽

A. Kroten ◽

M. Wojtkowska ◽

E. Oldak

Keyword(s):

Cerebrospinal Fluid ◽

Aseptic Meningitis ◽

Mumps Virus ◽

Echovirus 30

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Rapid Shell Vial Culture Technique for Detection of Enteroviruses and Adenoviruses in Fecal Specimens: Comparison with Conventional Virus Isolation Method

Journal of Clinical Microbiology ◽

10.1128/jcm.36.10.2865-2868.1998 ◽

1998 ◽

Vol 36 (10) ◽

pp. 2865-2868 ◽

Cited By ~ 33

Author(s):

G. J. J. Van Doornum ◽

J. C. De Jong

Keyword(s):

Cell Culture ◽

Cerebrospinal Fluid ◽

Virus Isolation ◽

Adenovirus Type ◽

Isolation Procedure ◽

Enzyme Linked Immunosorbent Assay ◽

Shell Vial ◽

Clinical Specimens ◽

Conventional Culture ◽

Stool Specimens

Detection of enteroviruses and adenoviruses mainly in fecal specimens by rapid culture with inoculation onto cell monolayers in flat-bottom tubes by centrifugation and immunofluorescence staining with genus-specific monoclonal antibodies was compared with that by the conventional virus isolation procedure. For both conventional culture and shell vial culture human lung fibroblast cells and tertiary monkey kidney cells were used. For enterovirus detection, 979 clinical specimens (916 stool specimens, 56 cerebrospinal fluid specimens, and 7 nasopharyngeal swabs) were used. Conventional culture detected 74 enterovirus isolates. A cytopathic effect compatible with the presence of an enterovirus after 3 days of incubation occurred in 25 of the 74 (34%) specimens that eventually became positive. The detection rate for enteroviruses by rapid cell culture after 2 to 3 days of incubation was 42 of 74 (57%). The genus-specific enterovirus monoclonal antibody did not react with strains of echovirus types 22 and 23 or enterovirus type 71. Rapid cell culture for the detection of adenoviruses was performed with 567 clinical specimens (536 stool specimens, 25 cerebrospinal fluid specimens, and 6 miscellaneous specimens), in which 42 adenoviruses were found by conventional culture. Nine of the 42 (21%) adenovirus isolates were detected by conventional culture within 3 days after inoculation, whereas 21 (50%) were found by rapid cell culture within 2 to 3 days. Only two of the nine specimens found to be positive for the enteric adenovirus type 41 by conventional culture as well by a type-specific enzyme-linked immunosorbent assay (ELISA) tested positive by rapid cell culture. In conclusion, the rapid shell vial assay allows the early detection and identification of enteroviruses and adenoviruses in clinical specimens but is markedly less sensitive than the conventional isolation procedure according to the eventual results of the conventional isolation procedure. Conventional cell culture remains a prerequisite for serotyping of enteroviral isolates. On the basis of the results for adenovirus type 41, the rapid detection of adenoviruses was not considered to be useful for the detection of clinically relevant adenoviruses in fecal samples.

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CLINICAL-EPIDEMIOLOGICAL CHARACTERISTICS OF ASEPTIC MENINGITIS IN CHILDREN OF KHMELNITSKYI REGION (PODILSKYI REGION, UKRAINE): FOURTEEN-YEAR EPIDEMIOLOGICAL OBSERVATION

Journal Infectology ◽

10.22625/2072-6732-2019-11-1-41-45 ◽

2019 ◽

Vol 11 (1) ◽

pp. 41-45

Author(s):

L. V. Pypa ◽

R. V. Svistilnik ◽

Yu. N. Lysytsia ◽

K. Yu. Romanchuk ◽

I. V. Odarchuk

Keyword(s):

Central Nervous System ◽

Cerebrospinal Fluid ◽

Nervous System ◽

Aseptic Meningitis ◽

Clinical Picture ◽

Epileptic Seizures ◽

Complete Recovery ◽

The Central Nervous System ◽

Meningitis In Children ◽

Epidemiological Characteristics

Aim of work – to analyze the etiological structure, epidemiological structure, social-demographic features and the nature of the development of complications of the central nervous system in aseptic meningitis in children in Khmelnitskyi region for the period 2004-2017.Materials and methods. It was conducted a prospective analysis of 208 cases of aseptic meningitis in children of whom 138 people were boys and 70 people were girls. The etiology of the disease was determined by studying cerebrospinal fluid using PCR method. Complications of the central nervous system were diagnosed on the basis of the clinical picture and CT or MRI scans. The analytical method was used to conduct the analysis of the received data.Results. The highest seasonal increase of the incidence was from August to October and it was 65.6% with its peak in September (24.0%). The clinical picture was characterized by a moderate trend in 71.2% of cases and in 28.8% by a severe course. In 100% of cases the disease began with a fever, headache (83.6%), vomiting (76.9%), abdominal pain with diarrhea (6.2%), epileptic seizures (0.9%). The average level of cytosis was 269.4±196.7 cells/mm3with a predominance of lymphocytes and the average protein level in cerebrospinal fluid was 73 ± 36 mg/dl. The etiological factor was established in 18 (8.6%) patients.Conclusions. Enterovirus remains to be the main pathogen which was determined in 72.2% of cases. The second place was taken by herpes viruses (22.2% of cases), the third place was given to the mumps virus (5.6% of cases) (in etiologically verified cases). In most cases the disease ended in complete recovery but in 47 (22.6%) patients the complications were observed. The prevalence of aseptic meningitis among children in Khmelnitskyi region was 6.2 per 100,000 children, and males outnumbered females by a 2:1 ratio.

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