ABSTRACT
The 2012 West Nile virus (WNV) epidemic was the largest since 2003 and the North Texas region was the most heavily impacted. We conducted a serosurvey of blood donors from four counties in the Dallas-Fort Worth area to characterize the epidemic. Blood donor specimens collected in November 2012 were tested for WNV-specific antibodies. Donors positive for WNV-specific IgG, IgM, and neutralizing antibodies were considered to have been infected in 2012. This number was adjusted using a multi-step process that accounted for timing of IgM seroreversion determined from previous longitudinal studies of WNV-infected donors. Of 4971 donations screened, 139 (2·8%) were confirmed WNV IgG positive, and 69 (1·4%) had IgM indicating infection in 2012. After adjusting for timing of sampling and potential seroreversion, we estimated that 1·8% [95% confidence interval (CI) 1·5-2·2] of the adult population in the Dallas-Fort Worth area were infected during 2012. The resulting overall estimate for the ratio of infections to reported WNV neuroinvasive disease (WNND) cases was 238:1 (95% CI 192-290), with significantly increased risk of WNND in older age groups. These findings were very similar to previous estimates of infections per WNND case, indicating no change in virulence as WNV evolved into an endemic infection in the United States.
Subject(s)
Epidemics , West Nile Fever/epidemiology , West Nile virus/physiology , Adolescent , Adult , Aged , Aged, 80 and over , Antibodies, Neutralizing/metabolism , Blood Donors/statistics & numerical data , Female , Humans , Immunoglobulin G/metabolism , Immunoglobulin M/metabolism , Incidence , Male , Middle Aged , Seroepidemiologic Studies , Texas/epidemiology , West Nile Fever/blood , West Nile Fever/virology , Young AdultABSTRACT
BACKGROUND: The US West Nile virus (WNV) epidemic in the summer and fall of 2002 included the first documented cases of transfusion-transmitted WNV infection. In December 2002, the FDA supported a voluntary market withdrawal by the blood banking community of frozen blood components collected in WNV high-activity areas. At the time, the prevalence of viremia and serologic markers for WNV in the blood supply was undefined. STUDY DESIGN AND METHODS: In collaboration with America's Blood Centers, 1468 frozen plasma components (of approx. 60,000 frozen units voluntarily withdrawn from the market) were selectively retrieved from the peak epidemic regions and season (June 23, 2002-September 28, 2002). These units were unlinked, subaliquoted, and tested by WNV enzyme immunoassays (EIAs; Focus Technologies and Abbott Laboratories) and nucleic acid amplification tests (NATs; Gen-Probe Inc. and Roche Molecular Systems). RESULTS: Of the 1468 EIA results from Abbott and Focus, 7 were anti-immunoglobulin M (IgM)- and anti-immunoglobulin G (IgG)-reactive by both assays, 8 and 1 were IgM-only-reactive, and 8 and 23 were IgG-only-reactive, respectively. NAT by Gen-Probe and Roche Molecular Systems yielded one RNA-positive, antibody-negative unit containing approximately 440 RNA copies per mL. An additional 10-fold replicate NAT testing by Gen-Probe on 14 of 15 IgM-reactive specimens yielded 2 additional IgM- and IgG-reactive units with low-level viremia (i.e., 7/10 and 2/10 replicates tested reactive). CONCLUSION: The prevalence of acute (RNA-positive) and recent (IgM-seroreactive) WNV infections indicates that transfusion risk in high-risk areas could have been considerable and that voluntary market withdrawal of frozen components likely averted some WNV transfusion transmissions. The existence of very-low-level viremic units raises concerns, because WNV minipool NAT screening will miss such units and individual NAT may not completely correct this situation.
Subject(s)
Blood Banks , Plasma/virology , West Nile Fever/blood , West Nile Fever/epidemiology , West Nile virus/isolation & purification , Antibodies, Viral/blood , Consumer Product Safety , Disease Outbreaks , Humans , Incidence , RNA, Viral/analysis , Risk Factors , Seroepidemiologic Studies , West Nile Fever/immunology , West Nile virus/genetics , West Nile virus/immunologyABSTRACT
In late summer 1999, the first domestically acquired human cases of WN encephalitis were documented in the USA. Aggressive vector-control and public education efforts by state and local public health officials limited the extent of human involvement. The discovery of virus-infected, overwintering mosquitoes during the winter of 1999-2000, predicted renewed virus activity for the following spring, and prompted early season vector-control activities and disease surveillance efforts in NYC and the surrounding areas. These surveillance efforts were focused on identifying WN virus infections in birds and mosquitoes as predictors of the potential risk of transmission to humans. By the end of the 2000 mosquito-borne disease transmission season, WN virus activity had been documented as far north as the states of Vermont and New Hampshire, and as far south as the state of North Carolina. The ongoing impacts that WN virus will have on wildlife, domestic animal and human populations of the western hemisphere are not yet known. Plans are in place for public health officials and scientists to monitor the further expansion of WN virus with the establishment or enhancement of vector-borne disease surveillance and control programs throughout the eastern seaboard. The valuable lessons learned from the detection and response to the introduction of WN virus into NYC should prove useful if and when subsequent intrusions of new disease agents occur.
Subject(s)
West Nile Fever/epidemiology , West Nile virus/isolation & purification , Animals , Disease Outbreaks , Ecosystem , Flavivirus/isolation & purification , Humans , Insect Vectors , New York City/epidemiology , North America/epidemiology , Population Surveillance , West Nile Fever/etiology , West Nile virus/geneticsABSTRACT
The development and application of nucleic acid sequence-based amplification (NASBA) assays for the detection of West Nile (WN) and St. Louis encephalitis (SLE) viruses are reported. Two unique detection formats were developed for the NASBA assays: a postamplification detection step with a virus-specific internal capture probe and electrochemiluminescence (NASBA-ECL assay) and a real-time assay with 6-carboxyfluorescein-labeled virus-specific molecular beacon probes (NASBA-beacon assay). The sensitivities and specificities of these NASBA assays were compared to those of a newly described standard reverse transcription (RT)-PCR and TaqMan assays for SLE virus and to a previously published TaqMan assay for WN virus. The NASBA assays demonstrated exceptional sensitivities and specificities compared to those of virus isolation, the TaqMan assays, and standard RT-PCR, with the NASBA-beacon assay yielding results in less than 1 h. These assays should be of utility in the diagnostic laboratory to complement existing diagnostic testing methodologies and as a tool in conducting flavivirus surveillance in the United States.
Subject(s)
Encephalitis Virus, St. Louis/isolation & purification , Encephalitis, St. Louis/virology , Self-Sustained Sequence Replication/methods , West Nile Fever/virology , West Nile virus/isolation & purification , Animals , Bird Diseases/virology , Birds/virology , Chlorocebus aethiops , Culicidae/virology , Encephalitis Virus, St. Louis/genetics , Molecular Probes , RNA, Viral/analysis , Reverse Transcriptase Polymerase Chain Reaction , Sensitivity and Specificity , Taq Polymerase/metabolism , Time Factors , Vero Cells , West Nile Fever/veterinaryABSTRACT
Widespread deaths of American Crows (Corvus brachyrhynchos)were associated with the 1999 outbreak of West Nile (WN) virus in the New York City region. We compared six organs from 20 crow carcasses as targets for WN virus detection. Half the carcasses had at least one positive test result for WN virus infection. The brain was the most sensitive test organ; it was the only positive organ for three of the positive crows. The sensitivity of crow organs as targets for WN virus detection makes crow death useful for WN virus surveillance.
Subject(s)
Bird Diseases/virology , Disease Outbreaks , Songbirds/virology , West Nile Fever/veterinary , West Nile virus/isolation & purification , Bird Diseases/pathology , New Jersey/epidemiology , RNA, Viral/analysis , Reverse Transcriptase Polymerase Chain Reaction , West Nile Fever/pathology , West Nile Fever/virology , West Nile virus/geneticsABSTRACT
An outbreak of encephalitis due to West Nile (WN) virus occurred in New York City and the surrounding areas during 1999. Mosquitoes were collected as part of a comprehensive surveillance program implemented to monitor the outbreak. More than 32,000 mosquitoes representing 24 species were tested, and 15 WN virus isolates were obtained. Molecular techniques were used to identify the species represented in the WN virus-positive mosquito pools. Most isolates were from pools containing Culex pipiens mosquitoes, but several pools contained two or more Culex species.
Subject(s)
Culex/virology , Disease Outbreaks , Insect Vectors/virology , West Nile virus/isolation & purification , Aedes/classification , Aedes/virology , Animals , Anopheles/classification , Anopheles/virology , Chlorocebus aethiops , Culex/classification , Culicidae/classification , Culicidae/virology , DNA, Viral/analysis , Insect Vectors/classification , New Jersey/epidemiology , New York/epidemiology , Vero Cells , West Nile Fever/epidemiology , West Nile Fever/virology , West Nile virus/geneticsABSTRACT
We describe two cases of West Nile (WN) encephalitis in a married couple in Tel Aviv, Israel, in 1999. Reverse transcription-polymerase chain reaction performed on a brain specimen from the husband detected a WN viral strain nearly identical to avian strains recovered in Israel in 1998 (99.9% genomic sequence homology) and in New York in 1999 (99.8%). This result supports the hypothesis that the 1999 WN virus epidemic in the United States originated from the introduction of a strain that had been circulating in Israel.
Subject(s)
West Nile Fever/virology , West Nile virus/genetics , Aged , Antibodies, Viral/blood , Antibodies, Viral/cerebrospinal fluid , Brain/virology , Female , Humans , Immunoglobulin M/blood , Immunoglobulin M/cerebrospinal fluid , Israel , Male , New York/epidemiology , RNA, Viral/analysis , Reverse Transcriptase Polymerase Chain Reaction , Treatment Outcome , West Nile Fever/blood , West Nile Fever/cerebrospinal fluid , West Nile Fever/epidemiology , West Nile Fever/immunology , West Nile virus/classification , West Nile virus/immunology , West Nile virus/isolation & purificationABSTRACT
West Nile (WN) virus was identified in the Western Hemisphere in 1999. Along with human encephalitis cases, 20 equine cases of WN virus were detected in 1999 and 23 equine cases in 2000 in New York. During both years, the equine cases occurred after human cases in New York had been identified.
Subject(s)
Disease Outbreaks , Horse Diseases/epidemiology , West Nile Fever/veterinary , West Nile virus/isolation & purification , Aedes/virology , Animals , Antibodies, Viral/analysis , Culex/virology , Horse Diseases/pathology , Horse Diseases/physiopathology , Horse Diseases/virology , Horses , Humans , New York/epidemiology , RNA, Viral/analysis , Reverse Transcriptase Polymerase Chain Reaction , West Nile Fever/epidemiology , West Nile Fever/pathology , West Nile Fever/physiopathology , West Nile virus/genetics , West Nile virus/immunologyABSTRACT
After the 1999 West Nile (WN) encephalitis outbreak in New York, 2,300 overwintering adult mosquitoes were tested for WN virus by cell culture and reverse transcriptase-polymerase chain reaction. WN viral RNA and live virus were found in pools of Culex mosquitoes. Persistence in overwintering Cx. pipiens may be important in the maintenance of WN virus in the northeastern United States.
Subject(s)
Culex/virology , Disease Outbreaks , Insect Vectors/virology , West Nile virus/isolation & purification , Aedes/cytology , Animals , Cell Line , Chlorocebus aethiops , New York City/epidemiology , RNA, Viral/analysis , Reverse Transcriptase Polymerase Chain Reaction , Seasons , Vero Cells , West Nile virus/geneticsABSTRACT
From July 25 to October 1, 1999, 826 patients were admitted to Volgograd Region, Russia, hospitals with acute aseptic meningoencephalitis, meningitis, or fever consistent with arboviral infection. Of 84 cases of meningoencephalitis, 40 were fatal. Fourteen brain specimens were positive in reverse transcriptase-polymerase chain reaction assays, confirming the presence of West Nile/Kunjin virus.
Subject(s)
Disease Outbreaks , West Nile Fever/epidemiology , Animals , Humans , Reverse Transcriptase Polymerase Chain Reaction , Russia/epidemiology , Time Factors , West Nile Fever/diagnosis , West Nile virus/classificationABSTRACT
The authors report on the development and application of a rapid TaqMan assay for the detection of West Nile (WN) virus in a variety of human clinical specimens and field-collected specimens. Oligonucleotide primers and FAM- and TAMRA-labeled WN virus-specific probes were designed by using the nucleotide sequence of the New York 1999 WN virus isolate. The TaqMan assay was compared to a traditional reverse transcriptase (RT)-PCR assay and to virus isolation in Vero cells with a large number ( approximately 500) of specimens obtained from humans (serum, cerebrospinal fluid, and brain tissue), field-collected mosquitoes, and avian tissue samples. The TaqMan assay was specific for WN virus and demonstrated a greater sensitivity than the traditional RT-PCR method and correctly identified WN virus in 100% of the culture-positive mosquito pools and 98% of the culture-positive avian tissue samples. The assay should be of utility in the diagnostic laboratory to complement existing human diagnostic testing and as a tool to conduct WN virus surveillance in the United States.
Subject(s)
Bird Diseases/diagnosis , Culicidae/virology , Reverse Transcriptase Polymerase Chain Reaction , Taq Polymerase/metabolism , West Nile Fever/diagnosis , West Nile virus/isolation & purification , Animals , Bird Diseases/virology , Birds/virology , Brain/virology , Chlorocebus aethiops , Humans , RNA, Viral/blood , RNA, Viral/cerebrospinal fluid , Sensitivity and Specificity , Vero Cells , Virus Cultivation , West Nile Fever/veterinary , West Nile Fever/virology , West Nile virus/geneticsABSTRACT
West Nile fever caused fatal disease in humans, horses, and birds in the northeastern United States during 1999. We studied birds from two wildlife facilities in New York City, New York, that died or were euthanatized and were suspected to have West Nile virus infections. Using standard histologic and ultrastructural methods, virus isolation, immunohistochemistry, in situ hybridization and reverse-transcriptase polymerase chain reaction, we identified West Nile virus as the cause of clinical disease, severe pathologic changes, and death in 27 birds representing eight orders and 14 species. Virus was detected in 23/26 brains (88%), 24/ 25 hearts (96%), 15/18 spleens (83%), 14/20 livers (70%), 20/20 kidneys (100%), 10/13 adrenals (77%), 13/ 14 intestines (93%), 10/12 pancreata (83%), 5/12 lungs (42%), and 4/8 ovaries (50%) by one or more methods. Cellular targets included neurons and glial cells in the brain, spinal cord, and peripheral ganglia; myocardial fibers; macrophages and blood monocytes; renal tubular epithelium; adrenal cortical cells; pancreatic acinar cells and islet cells; intestinal crypt epithelium; oocytes; and fibroblasts and smooth muscle cells. Purkinje cells were especially targeted, except in crows and magpies. Gross hemorrhage of the brain, splenomegaly, meningoencephalitis, and myocarditis were the most prominent lesions. Immunohistochemistry was an efficient and reliable method for identifying infected cases, but the polyclonal antibody cross-reacted with St. Louis encephalitis virus and other flaviviruses. In contrast, the in situ hybridization probe pWNV-E (WN-USAMRIID99) reacted only with West Nile virus. These methods should aid diagnosticians faced with the emergence of West Nile virus in the United States.
Subject(s)
Bird Diseases/pathology , Disease Outbreaks/veterinary , West Nile Fever/veterinary , Animals , Birds , Immunohistochemistry , In Situ Hybridization/veterinary , Microscopy, Electron/veterinary , New York City , West Nile Fever/pathology , West Nile virusABSTRACT
West Nile virus is a mosquito borne flavivirus endemic over a large geographic area including Africa, Asia, and the Middle East. Although the virus generally causes a mild, self-limiting febrile illness in humans, it has sporadically caused central nervous system infections during epidemics. An isolate of West Nile virus was obtained from a pool of four male Culex univittatus complex mosquitoes while we were conducting an investigation of Rift Valley fever along the Kenya-Uganda border in February-March 1998. This represents the first field isolation of West Nile virus from male mosquitoes and strongly suggests that vertical transmission of the virus occurs in the primary maintenance mosquito vector in Kenya. A phylogenetic analysis of the complete amino acid sequence of the viral envelope glycoprotein demonstrated a sister relationship with a Culex pipiens mosquito isolate from Romania made in 1996. This unexpected finding probably reflects the role of migratory birds in disseminating West Nile virus between Africa and Europe.
Subject(s)
Culex/virology , Infectious Disease Transmission, Vertical , Insect Vectors/virology , West Nile Fever/transmission , West Nile virus/immunology , Amino Acid Sequence , Animals , Antigens, Viral/analysis , Base Sequence , Chlorocebus aethiops , DNA Primers/chemistry , DNA, Viral/chemistry , Electrophoresis, Agar Gel , Female , Fluorescent Antibody Technique, Indirect , Humans , Kenya/epidemiology , Molecular Sequence Data , Phylogeny , RNA, Viral/chemistry , RNA, Viral/isolation & purification , Reverse Transcriptase Polymerase Chain Reaction , Sequence Alignment , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Vero Cells , Viral Envelope Proteins/chemistry , Viral Envelope Proteins/genetics , West Nile Fever/epidemiology , West Nile Fever/immunology , West Nile Fever/virology , West Nile virus/classification , West Nile virus/geneticsABSTRACT
In late summer 1999, an outbreak of human encephalitis occurred in the northeastern United States that was concurrent with extensive mortality in crows (Corvus species) as well as the deaths of several exotic birds at a zoological park in the same area. Complete genome sequencing of a flavivirus isolated from the brain of a dead Chilean flamingo (Phoenicopterus chilensis), together with partial sequence analysis of envelope glycoprotein (E-glycoprotein) genes amplified from several other species including mosquitoes and two fatal human cases, revealed that West Nile (WN) virus circulated in natural transmission cycles and was responsible for the human disease. Antigenic mapping with E-glycoprotein-specific monoclonal antibodies and E-glycoprotein phylogenetic analysis confirmed these viruses as WN. This North American WN virus was most closely related to a WN virus isolated from a dead goose in Israel in 1998.
Subject(s)
Disease Outbreaks , West Nile Fever/epidemiology , West Nile Fever/virology , West Nile virus/classification , West Nile virus/genetics , Amino Acid Sequence , Animals , Antibodies, Monoclonal/immunology , Antibodies, Viral/immunology , Base Sequence , Bird Diseases/epidemiology , Bird Diseases/virology , Birds/virology , Encephalitis Viruses, Japanese/classification , Encephalitis Viruses, Japanese/genetics , Fluorescent Antibody Technique, Indirect , Genome, Viral , Humans , Molecular Sequence Data , New England/epidemiology , New York City/epidemiology , Phylogeny , Songbirds/virology , Viral Envelope Proteins/chemistry , Viral Envelope Proteins/genetics , Viral Envelope Proteins/immunology , West Nile Fever/veterinary , West Nile virus/immunology , West Nile virus/isolation & purificationABSTRACT
O'nyong-nyong (ONN) fever, caused by infection with a mosquito-borne central African alphavirus, is an acute, nonfatal illness characterized by polyarthralgia. During 1996-1997, south-central Uganda experienced the second ONN fever epidemic ever recognized. Among 391 persons interviewed and sampled, 40 cases of confirmed and 21 of presumptive, well-characterized acute, recent, or previous ONN fever were identified through active case-finding efforts or during a household serosurvey and by the application of clinical and laboratory criteria. Among confirmed cases, the knees and ankles were the joints most commonly affected. The median duration of arthralgia was 6 days (range, 2-21 days) and of immobilization was 4 days (range, 1-14 days). In the majority, generalized skin rash was reported, and nearly half had lymphadenopathy, mainly of the cervical region. Viremia was documented in 16 cases, primarily during the first 3 days of illness, and in some of these, body temperature was normal. During this epidemic, the combination of fever, arthralgia, and lymphadenopathy had a specificity of 83% and a sensitivity of 61% in the identification of cases of ONN fever and thus could be useful for surveillance purposes.
Subject(s)
Alphavirus Infections/epidemiology , Arthralgia/epidemiology , Fever/epidemiology , Lymphatic Diseases/epidemiology , Adolescent , Adult , Aged , Aged, 80 and over , Alphavirus Infections/diagnosis , Antibodies, Viral/blood , Child , Child, Preschool , Female , Humans , Immunoglobulin M/blood , Male , Middle AgedABSTRACT
O'nyong-nyong (ONN) virus is an alphavirus (family Togaviridae, genus Alphavirus) classified in the Semliki Forest virus (SFV) antigenic complex. ONN was initially isolated in northern Uganda in 1959 during the early stages of an explosive arbovirus epidemic in which > 2 million cases were reported. No additional epidemics or human isolations of ONN were reported until 1996, when it was isolated from an epidemic in southern Uganda. We report the complete nucleotide and deduced amino acid sequence of one of these 1996-1997 ONN isolates (SG650) and that of the related alphavirus Igbo Ora virus. The data indicate that the recent ONN virus isolate is closely related to the previously published ONN strain isolated in 1959. In addition, phylogenetic analysis of the sequence data reveals that Igbo Ora virus, previously thought to be a separate virus closely related to ONN and Chikungunya (CHIK), clearly is a strain of ONN. The sequence data also reveal that unlike the published ONN (1959) sequence, all ONN strains from the 1996-1997 epidemic possess a stop codon at the nsp3-nsp4 junction.
Subject(s)
Alphavirus Infections/epidemiology , Alphavirus Infections/virology , Chikungunya virus/genetics , Genome, Viral , Adolescent , Amino Acid Sequence , Animals , Base Sequence , Chikungunya virus/classification , Chlorocebus aethiops , Culicidae , Humans , Insect Vectors , Male , Molecular Sequence Data , Phylogeny , Uganda/epidemiology , Vero CellsABSTRACT
Nucleotide sequences of the envelope protein genes of 19 geographically and temporally distinct dengue (DEN)-4 viruses were determined. Nucleic acid sequence comparison revealed that the identity among the DEN-4 viruses was greater than 92%. Similarity among deduced amino acids was between 96 and 100%; in most cases identical amino acid substitutions occurred among viruses from similar geographical regions. Alignment of nucleic acid sequences followed by parsimony analysis generated phylogenetic trees, which indicated that geographically independent evolution of DEN-4 viruses had occurred. DEN-4 viruses were separated into two genetically distinct subtypes (genotypes). Genotype-1 contains viruses from the Philippines, Thailand and Sri Lanka; genotype-2 consists of viruses from Indonesia, Tahiti, the Caribbean Islands (Puerto Rico, Dominica) and Central and South America.
Subject(s)
Dengue Virus/genetics , Evolution, Molecular , Phylogeny , Amino Acid Sequence , Animals , Culicidae/virology , Genes, Viral/genetics , Genotype , Humans , Molecular Sequence Data , Neutralization Tests , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Sequence Homology, Nucleic Acid , Viral Envelope Proteins/genetics , Viral Structural Proteins/geneticsABSTRACT
Colorado tick fever (CTF) virus elicits an acute illness in humans, producing nonspecific flu-like symptoms and a biphasic fever in approximately 50% of patients. The disease is transmitted by the adult Rocky Mountain wood tick (Dermacentor andersoni), and therefore incidence is limited by the habitat and life cycle of that vector. The early symptoms of infection are difficult to distinguish from those of several other agents, especially Rickettsia rickettsii. Serologic testing is usually unable to provide evidence of CTF viral infection during the acute phase because of the late appearance of the various antibodies. Here we report the development and clinical application of a test to diagnose this disease during the acute stages. Oligonucleotide primers to the S2 segment of CTF (Florio) virus were made, and these were used in the amplification of a 528-bp fragment of DNA, transcribed from the double-stranded CTF virus RNA template by reverse transcriptase PCR. RNAs processed from 16 CTF virus isolates yielded similar results when analyzed on agarose gels. These were distinguishable from their antigenic relatives Eyach, S6-14-03, and T5-2092 and from other coltiviruses and an orbivirus but not from the antigenically distinct CTF virus-related isolate 720896. A mouse model demonstrated the utility of this method with whole-blood specimens, and CTF virus was successfully detected in human sera from the initial day of the onset of symptoms to 8 days later. The reverse transcriptase PCR method is a promising tool for the early diagnosis of CTF viral infection, or for ruling out CTF virus as the etiologic agent, in order to facilitate appropriate medical support.