Detection of West Nile Virus by the Polymerase Chain Reaction and Analysis of Nucleotide Sequence Variation

Porter, Kevin R.; Summers, Peter L.; Dubois, Doria R.; Puri, Beena; Nelson, William M.; Henchal, Erik A.; Oprandy, John J.; Hayes, Curtis G.

doi:10.4269/ajtmh.1993.48.440

Cited by 36 publications

(23 citation statements)

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“…Similarly, Porter et al (17) reported that most mutations among seven WN virus isolates from six countries were silent. However, Chambers et al (20) reported a single nucleotide substitution of T to A at position 463 of the WN virus envelope gene that specified the substitution of asparagine for tyrosine at amino acid site 155 and produced an N-linked glycosylation site.…”

Section: Discussionmentioning

confidence: 99%

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A phylogenetic approach to following West Nile virus in Connecticut

Anderson

Vossbrinck

Andreadis

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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Although 34 of the isolates showed sequences identical to the WN-NY99 isolate, we were able to show geographical-based clusters of mutations. In particular, 26 isolates were characterized by mutation of C to T at position 858. This group apparently originated in Stamford, CT and disseminated to sites located as far as 54 miles from Stamford. Sequences of WN virus isolated from both brain and heart tissues from the same avian host were identical in all 14 tested individual birds, suggesting that the mutations we have documented are real and not caused by culture, RNA extraction, or PCR procedures. We conclude that this portion of the viral genome will enable us to follow the geographical and temporal movement of variant WN virus strains as they adapt to North America. viral evolution ͉ epidemiology

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Section: Discussionmentioning

confidence: 99%

“…Extensive genetic variation exists among WN virus isolates in Africa, Australia, Asia, and Europe (15)(16)(17). Phylogenetic studies of isolates from Europe and Africa suggest the introduction of WN virus into Europe by birds migrating out of sub-Saharan Africa.…”

mentioning

confidence: 99%

A phylogenetic approach to following West Nile virus in Connecticut

Anderson

Vossbrinck

Andreadis

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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“…Although the serum samples were not transported under appropriate temperature conditions, an attempt was made to isolate virus from the serum samples using methods available for isolation including mouse and cell culture inoculations. 20 …”

Section: Methodsmentioning

confidence: 99%

An outbreak of West Nile fever among migrants in Kisangani, Democratic Republic of Congo.

Nur¹,

Groen²,

Heuvelmans³

et al. 1999

The American Journal of Tropical Medicine and Hygiene

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“…Such assays should enhance surveillance effectiveness both by increasing the number of laboratories capable of performing tests and by lengthening the period in which vector control can be undertaken (4,12,28,29,31). Within the last 10 years, rapid, sensitive and specific nucleic acid-based assays employing RT-PCR or TaqMan have been developed for several RNA viruses, including WN virus (17,22). In connection with developing an ELISA-based Ag detection assay for WN virus, we have also compared the performance of this test with TaqMan and traditional RT-PCR using two blindly coded sets of field-collected specimens.…”

Section: Discussionmentioning

confidence: 99%

Detection of West Nile Virus Antigen in Mosquitoes and Avian Tissues by a Monoclonal Antibody-Based Capture Enzyme Immunoassay

et al. 2002

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An antigen capture immunoassay to detect West Nile (WN) virus antigen in infected mosquitoes and avian tissues has been developed. With this assay purified WN virus was detected at a concentration of 32 pg/0.1 ml, and antigen in infected suckling mouse brain and laboratory-infected mosquito pools could be detected when the WN virus titer was 10 2.1 to 10 3.7 PFU/0.1 ml. In a blindly coded set of field-collected mosquito pools (n ‫؍‬ 100), this assay detected WN virus antigen in 12 of 18 (66.7%) TaqMan-positive pools, whereas traditional reverse transcriptase PCR detected 10 of 18 (55.5%) positive pools. A sample set of 73 organ homogenates from naturally infected American crows was also examined by WN virus antigen capture immunoassay and TaqMan for the presence of WN virus. The antigen capture assay detected antigen in 30 of 34 (88.2%) TaqMan-positive tissues. Based upon a TaqMan-generated standard curve of infectious WN virus, the limit of detection in the antigen capture assay for avian tissue homogenates was approximately 10 3 PFU/0.1 ml. The recommended WN virus antigen capture protocol, which includes a capture assay followed by a confirmatory inhibition assay used to retest presumptive positive samples, could distinguish between the closely related WN and St. Louis encephalitis viruses in virus-infected mosquito pools and avian tissues. Therefore, this immunoassay demonstrates adequate sensitivity and specificity for surveillance of WN virus activity in mosquito vectors and avian hosts, and, in addition, it is easy to perform and relatively inexpensive compared with the TaqMan assay.

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Detection of West Nile Virus by the Polymerase Chain Reaction and Analysis of Nucleotide Sequence Variation

Cited by 36 publications

References 0 publications

A phylogenetic approach to following West Nile virus in Connecticut

A phylogenetic approach to following West Nile virus in Connecticut

An outbreak of West Nile fever among migrants in Kisangani, Democratic Republic of Congo.

Detection of West Nile Virus Antigen in Mosquitoes and Avian Tissues by a Monoclonal Antibody-Based Capture Enzyme Immunoassay

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