Solid-phase immune electron microscopy-double-antibody technique for rapid detection of papovaviruses

Giraldo, Gaetano; Beth, E.; Lee, J; Harven, E D; Chernesky, Max

doi:10.1128/jcm.15.3.517-521.1982

Cited by 14 publications

(3 citation statements)

References 21 publications

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“…Using SPIEM, the sensitivity of diagnostic EM is improved up to 100 times (Kjeldsberg and Mortensen-Egnund, 1982) and reaches detection limits down to values of e.g. 10 2 -10 3 plaque forming units per milliliter (Giraldo et al, 1982). Compared to nucleic acid amplification techniques (NAT) for SRSVs neither SPIEM nor NAT detect every virus-positive specimen-however, they complement each other (Humphrey et al, 1997;Cubitt et al, 1999).…”

Section: Methods Used In Diagnostic Emmentioning

confidence: 99%

Diagnostic electron microscopy is still a timely and rewarding method

Biel

Gelderblom

1999

Journal of Clinical Virology

116

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In the history of diagnostic EM in virology, a change in use has been seen. Starting in the 1990s and coincident with the broad introduction of 'modern' diagnostic techniques, the number of EM diagnostic labs has decreased considerably--in spite of the obvious advantages of this technique. To guarantee the continuing performance of diagnostic EM in the future. EQA runs have to be performed as with other techniques in the diagnostic armament. The growing number of participants and participating countries indicates an interest in as well as a need for this program.

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Section: Methods Used In Diagnostic Emmentioning

confidence: 99%

Diagnostic electron microscopy is still a timely and rewarding method

Biel

Gelderblom

1999

Journal of Clinical Virology

116

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“…Traditional methods for detecting and identifying polyomaviruses include serologic methods, virus isolation by cell culture and electron microscopy [Giraldo et al, 1982;Knowles et al, 1989;Nilsen et al, 1991]. Recently, several studies have shown PCR to be an effective tool for detecting polyomaviruses in a range of clinical samples Chang et al, 1996;De Santis and Azzi, 1996]; as a result, PCR is now emerging as the ''gold standard'' for polyomavirus detection.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous detection and differentiation of human polyomaviruses JC and BK by a rapid and sensitive PCR‐ELAHA assay and a survey of the JCV subtypes within an Australian population

Whiley

Arden

Mackay

et al. 2004

Journal of Medical Virology

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Human polyomaviruses JCV and BKV can cause several clinical manifestations in immunocompromised hosts, including progressive multifocal leukoencephalopathy (PML) and haemorrhagic cystitis. Molecular detection by polymerase chain reaction (PCR) is recognised as a sensitive and specific method for detecting human polyomaviruses in clinical samples. In this study, we developed a PCR assay using a single primer pair to amplify a segment of the VP1 gene of JCV and BKV. An enzyme linked amplicon hybridisation assay (ELAHA) using species-specific biotinylated oligonucleotide probes was used to differentiate between JCV and BKV. This assay (VP1-PCR-ELAHA) was evaluated and compared to a PCR assay targeting the human polyomavirus T antigen gene (pol-PCR). DNA sequencing was used to confirm the polyomavirus species identified by the VP1-PCR-ELAHA and to determine the subtype of each JCV isolate. A total of 297 urine specimens were tested and human polyomavirus was detected in 105 specimens (35.4%) by both PCR assays. The differentiation of JCV and BKV by the VP1-PCR-ELAHA showed good agreement with the results of DNA sequencing. Further, DNA sequencing of the JCV positive specimens showed the most prevalent JCV subtype in our cohort was 2a (27%) followed by 1b (20%), 1a (15%), 2c (14%), 4 (14%) and 2b (10%). The results of this study show that the VP1-PCR-ELAHA is a sensitive, specific and rapid method for detecting and differentiating human polyomaviruses JC and BK and is highly suitable for routine use in the clinical laboratory.

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“…Others have used SPIEMDAT (SPIEMDAGT without colloidal gold) for the detection of rotavirus (9) and papovavirus (5). In the former study, grids were precoated with protein A to facilitate antibody binding (9), while in the latter study no decorator antibody was employed (5). Papovavirus was detected by using the same antiserum for the capture and decorator antibodies (5).…”

Section: Discussionmentioning

confidence: 99%

Sensitive solid-phase immune electron microscopy double-antibody technique with gold-immunoglobulin G complexes for detecting rotavirus in cell culture and feces

Wu¹,

Mahony²,

Simon³

et al. 1990

J Clin Microbiol

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A new solid-phase immune electron microscopy double-antibody colloidal-gold technique (SPIEMDAGT) was developed and compared with direct electron microscopy, direct immune electron microscopy, and enzyme immunoassay for detecting rotavirus. Guinea pig and rabbit antirotavirus antisera were used as capture and detector antibodies, respectively, and goat anti-rabbit immunoglobulin G-gold complexes were employed as a label. Animal rotavirus in cell culture media and human virus in stool specimens were detected by this method. On average, SPIEMDAGT detected 800 times more virus particles than direct electron microscopy and 45 times more particles than direct immune electron microscopy and yielded 20% more positives than enzyme immunoassay. SPIEMDAGT could detect not only viral antigen associated with morphologically recognizable particles but also antigen present when whole virus particles were not visible.

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Solid-phase immune electron microscopy-double-antibody technique for rapid detection of papovaviruses

Cited by 14 publications

References 21 publications

Diagnostic electron microscopy is still a timely and rewarding method

Diagnostic electron microscopy is still a timely and rewarding method

Simultaneous detection and differentiation of human polyomaviruses JC and BK by a rapid and sensitive PCR‐ELAHA assay and a survey of the JCV subtypes within an Australian population

Sensitive solid-phase immune electron microscopy double-antibody technique with gold-immunoglobulin G complexes for detecting rotavirus in cell culture and feces

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