RESUMO
During 2019-2020, the Virgin Islands Department of Health investigated potential animal reservoirs of Leptospira spp., the bacteria that cause leptospirosis. In this cross-sectional study, we investigated Leptospira spp. exposure and carriage in the small Indian mongoose (Urva auropunctata, syn: Herpestes auropunctatus), an invasive animal species. This study was conducted across the three main islands of the U.S. Virgin Islands (USVI), which are St. Croix, St. Thomas, and St. John. We used the microscopic agglutination test (MAT), fluorescent antibody test (FAT), real-time polymerase chain reaction (lipl32 rt-PCR), and bacterial culture to evaluate serum and kidney specimens and compared the sensitivity, specificity, positive predictive value, and negative predictive value of these laboratory methods. Mongooses (n = 274) were live-trapped at 31 field sites in ten regions across USVI and humanely euthanized for Leptospira spp. testing. Bacterial isolates were sequenced and evaluated for species and phylogenetic analysis using the ppk gene. Anti-Leptospira spp. antibodies were detected in 34% (87/256) of mongooses. Reactions were observed with the following serogroups: Sejroe, Icterohaemorrhagiae, Pyrogenes, Mini, Cynopteri, Australis, Hebdomadis, Autumnalis, Mankarso, Pomona, and Ballum. Of the kidney specimens examined, 5.8% (16/270) were FAT-positive, 10% (27/274) were culture-positive, and 12.4% (34/274) were positive by rt-PCR. Of the Leptospira spp. isolated from mongooses, 25 were L. borgpetersenii, one was L. interrogans, and one was L. kirschneri. Positive predictive values of FAT and rt-PCR testing for predicting successful isolation of Leptospira by culture were 88% and 65%, respectively. The isolation and identification of Leptospira spp. in mongooses highlights the potential role of mongooses as a wildlife reservoir of leptospirosis; mongooses could be a source of Leptospira spp. infections for other wildlife, domestic animals, and humans.
Assuntos
Reservatórios de Doenças/microbiologia , Herpestidae/microbiologia , Leptospira/isolamento & purificação , Testes de Aglutinação , Animais , Estudos Transversais , Herpestidae/fisiologia , Humanos , Espécies Introduzidas/estatística & dados numéricos , Rim/microbiologia , Leptospira/genética , Leptospira/imunologia , Leptospirose/microbiologia , Leptospirose/transmissão , Filogenia , Ilhas Virgens AmericanasRESUMO
The emergence and spread of Zika virus (ZIKV) presented a challenge to the diagnosis of ZIKV infections in areas with transmission of dengue (DENV) and chikungunya (CHIKV) viruses. To facilitate detection of ZIKV infections, and differentiate these infections from DENV and CHIKV, we developed the Trioplex real-time RT-PCR assay (Trioplex assay). Here, we describe the optimization of multiplex and singleplex formats of the assay for a variety of chemistries and instruments to facilitate global standardization and implementation. We evaluated the analytical performance of all Trioplex modalities for detection of these three pathogens in serum and whole blood, and for ZIKV in urine. The limit of detection for the three viruses and in different RNA-extraction modalities is near 103 genome copy equivalents per milliliter (GCE/mL). Simultaneous testing of more than one specimen type from each patient provides a 6.4% additional diagnostic sensitivity. Overall, the high sensitivity of the Trioplex assay demonstrates the utility of this assay ascertaining Zika cases.
Assuntos
Febre de Chikungunya/diagnóstico , Vírus Chikungunya/genética , Vírus da Dengue/genética , Dengue/diagnóstico , Reação em Cadeia da Polimerase Multiplex/métodos , Infecção por Zika virus/diagnóstico , Zika virus/genética , Calibragem , Febre de Chikungunya/sangue , Febre de Chikungunya/virologia , Vírus Chikungunya/isolamento & purificação , Dengue/sangue , Dengue/virologia , Vírus da Dengue/isolamento & purificação , Humanos , Limite de Detecção , Reação em Cadeia da Polimerase Multiplex/normas , Kit de Reagentes para Diagnóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/normas , Zika virus/isolamento & purificação , Infecção por Zika virus/urina , Infecção por Zika virus/virologiaRESUMO
The exotic arboviruses chikungunya (CHIKV) and Zika (ZIKV) recently caused large outbreaks and continue to circulate in Puerto Rico, prompting entomological investigations at 9 locations with confirmed CHIKV- or ZIKV-infected human cases. Adult mosquitoes were collected using the Centers for Disease Control and Prevention autocidal gravid ovitraps over a 14-day period at each site. Mean female Aedes aegypti captured per trap-week ranged from 13.47 per trap-week to 1.27 per trap-week. Arbovirus-positive pools were detected at 7 of the 9 sampling sites. We investigated vertical transmission by collecting Ae. aegypti eggs in a single location where ZIKV was found in adult mosquitoes. We discuss the relationship between vector density and infection rates and its implications for determining mosquito density thresholds of novel invasive arboviruses such as CHIKV and ZIKV.