An immune dot-blot technique for the diagnosis of ocular adenovirus infection

Killough, Rosemary; Klapper, Paul E.; Bailey, A. S.; Sharp, I. R.; Tullo, Andrew B.; Richmond, S J

doi:10.1016/0166-0934(90)90020-g

Cited by 9 publications

(3 citation statements)

References 14 publications

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“…The direct immunofluorescence test was less sensitive than PCR in our study. In addition, adeno-immunofluorescence tests have been shown to produce false positive results, probably due to cross-reactivity with contaminating staphylococcal protein A [16]. Tissue culture, the other major alternative to PCR and direct immunofluorescence, theoretically only requires a single infectious unit in the inoculum and is therefore considered highly sensitive.…”

Section: Discussionmentioning

confidence: 99%

“…Immunologic tests, on the other hand, are more rapid. However, they rely on the presence of large amounts of reactive adenoviral antigen in the samples [16]. Serologic tests usually utilize the 360-kD hexon trimer, a major adenovirion capsomere protein that characterizes some subgroup- and group-specific determinants common to all mammalian adenoviruses.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of Polymerase Chain Reaction for the Detection of Adenoviruses in Conjunctival Swab Specimens Using Degenerate Primers in Comparison with Direct Immunofluorescence

Weitgasser¹,

Haller²,

El-Shabrawi³

2002

Ophthalmologica

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Purpose: To investigate a rapid and sensitive polymerase chain reaction (PCR)-based assay for the detection of adenoviral infections in conjunctival swabs. Methods: Degenerate primers derived from the adenoviral hexon gene were designed for amplification of a 302-bp product by PCR. In 15 ocular swab specimens, using a simple sample preparation method, PCR was compared with a commercially available direct immunofluorescence test. Results: Fourteen samples of patients with clinically diagnosed adenoviral keratoconjunctivitis, when screened by direct immunofluorescence, were positive for adenoviral protein. In all samples positive by immunofluorescence, the 302-bp product was amplifiable. The sample negative by direct immunofluorescence was positive using PCR. Conclusion: PCR using degenerate primers proved to have several advantages over current diagnostic techniques. It offers considerable improvement in sensitivity over immunoassays and speed over tissue culture isolation and is a highly potential tool for the diagnosis of adenoviral ocular infections.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Evaluation of Polymerase Chain Reaction for the Detection of Adenoviruses in Conjunctival Swab Specimens Using Degenerate Primers in Comparison with Direct Immunofluorescence

Weitgasser¹,

Haller²,

El-Shabrawi³

2002

Ophthalmologica

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“…In 1991, 52 405 specimens were received, I1 255 for virus detection, 14 595 for chlamydia detection, and 26 555 for virus serology. Tests in routine use include virus isolation in cell culture, respiratory virus antigen detection by immunofluorescence (Morris and Semple, 1990b), cytomegalovirus immediate early antigen detection (Morris et al, 1987) the polymerase chain reaction for the diagnosis of herpes simplex virus encephalitis adenovirus immune dot-blot (Killough et al, 1990) chlamydial isolation in cell culture and immune dot-blot (Mearns et al, 1988) electron microscopy including serotyping of faecal adenoviruses (Wood et al, 1989) rotavirus enzyme-linked immunosorbent assay, hepatitis A, B, C and D virus and human immunodeticiency virus (HIV) serological tests (Morris et al, 1990a), rubella and toxoplasma screening and serological diagnosis, complement fixation tests, screening for recent B 19 parvovirus infection (Rayment et al, 1990), and cytomegalovirus antibody tests on organ donors (Morris et al, 1990~).…”

Section: Laboratorymentioning

confidence: 99%

Computer network for a diagnostic virology laboratory

Stokes

Morris

Klapper

et al. 1993

Journal of Virological Methods

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A data base for a large diagnostic virology laboratory is described. The system uses a network of personal computers. It allows the entry, long-term storage, and subsequent retrieval of specimen and patient records (comprising personal identifiers and specimen and result information), and hard-copy results reporting. Sited entirely within the laboratory, the network is not connected to a modem. Within the laboratory there is restricted access to human immunodeficiency virus test results to guarantee patient confidentiality. Retention of a hard-copy of specimen request cards ensures the availability of the original clinical information. The data base is copied on a second file server to facilitate searches, and daily streaming onto magnetic tape provides system protection in the event of hard disc failure. Matching of old and new patient records is done by surname, date of birth, and sex, and therefore duplicate records accumulate when patient names are misspelt on specimen request forms. The system requires further development to speed searches of the data base and to achieve automatic generation of laboratory worksheets. Future goals are the replacement of hard-copy records of clinical information and hard-copy reporting with on-line access to hospital data bases and on-line requesting by and reporting to the clinician.

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Comparison of primer sets for detection of fecal and ocular adenovirus infection using the polymerase chain reaction

et al. 1996

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Adenoviruses of subgenus F (types 40 and 41) cause infantile gastroenteritis and adenoviruses principally of types 1-7 are found in feces during respiratory or generalized infections. Adenoviruses (mostly types 3, 4, 8, 19, or 37) are also linked with follicular or epidemic conjunctivitis. The diagnostic efficiency of the polymerase chain reaction (PCR) for adenoviruses was assessed using genus-reactive primers H1 and H2 or JCH1 and JCH2 or subgenus F-specific primers F1a and F2a. With diarrheal stool specimens containing subgenus F adenoviruses, F1a/F2a PCR achieved at least as high a positivity rate (75/76 [99%]) as electron microscopy (72/76 [95%]) and was more sensitive than polyclonal antibody-based immune electron microscopy (IEM) for subgenus identification (75/76 [99%] vs. 66/76 [87%], P = 0.008). Twenty-three subgenus F strains untypeable by monoclonal antibody-based IEM were typed as 40 (n = 4) or 41 (n = 19) by Hha I digestion of the PCR product. The genus-reactive primer pairs provided DNA amplification assays of generally equal efficiency on conjunctival swab specimens though possible nucleic acid degradation in DNA extracts during storage could have meant that JCH1/JCH2 PCR was truly the more sensitive. The use of either genus-reactive primer set on fecal specimens cannot be recommended because, although the positivity rates with subgenus F PCR positive specimens were high (70/75 [93%] for H1 and H2, 14/15 [93%] for JCH1 and JCH2), the detection rates were disappointing with similar specimens yielding nonsubgenus F adenoviruses.

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An immune dot-blot technique for the diagnosis of ocular adenovirus infection

Cited by 9 publications

References 14 publications

Evaluation of Polymerase Chain Reaction for the Detection of Adenoviruses in Conjunctival Swab Specimens Using Degenerate Primers in Comparison with Direct Immunofluorescence

Evaluation of Polymerase Chain Reaction for the Detection of Adenoviruses in Conjunctival Swab Specimens Using Degenerate Primers in Comparison with Direct Immunofluorescence

Computer network for a diagnostic virology laboratory

Comparison of primer sets for detection of fecal and ocular adenovirus infection using the polymerase chain reaction

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