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OBJECTIVES: Leptospira, the spirochaete causing leptospirosis, can be classified into >250 antigenically distinct serovars. Although knowledge of the animal host species and geographic distribution of Leptospira serovars is critical to understand the human and animal epidemiology of leptospirosis, current data are fragmented. We aimed to systematically review, the literature on animal host species and geographic distribution of Leptospira serovars to examine associations between serovars with animal host species and regions and to identify geographic regions in need of study. METHODS: Nine library databases were searched from inception through 9 March 2023 using keywords including Leptospira, animal, and a list of serovars. We sought reports of detection of Leptospira, from any animal, characterised by cross agglutinin absorption test, monoclonal antibody typing, serum factor analysis, or pulsed-field gel electrophoresis to identify the serovar. RESULTS: We included 409 reports, published from 1927 through 2022, yielding data on 154 Leptospira serovars. The reports included data from 66 (26.5%) of 249 countries. Detections were from 144 animal host species including 135 (93.8%) from the class Mammalia, 5 (3.5%) from Amphibia, 3 (2.1%) from Reptilia, and 1 (0.7%) from Arachnida. Across the animal host species, Leptospira serovars that were detected in the largest number of animal species included Grippotyphosa (n = 39), Icterohaemorrhagiae (n = 29), Pomona (n = 28), Australis (n = 25), and Ballum (n = 25). Of serovars, 76 were detected in a single animal host species. We created an online database to identify animal host species for each serovar by country. CONCLUSIONS: We found that many countries have few or no Leptospira serovars detected from animal host species and that many serovars were detected from a single animal species. Our study highlights the importance of efforts to identify animal host species of leptospirosis, especially in places with a high incidence of human leptospirosis. We provide an updated resource for leptospirosis researchers.
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Leptospira , Leptospirose , Sorogrupo , Leptospira/classificação , Animais , Leptospirose/epidemiologia , Leptospirose/microbiologia , HumanosRESUMO
Objectives: Leptospira, the spirochaete causing leptospirosis, can be classified into >250 antigenically distinct serovars. Although knowledge of the animal host species and geographic distribution of Leptospira serovars is critical to understand the human and animal epidemiology of leptospirosis, currently data are fragmented. We aimed to systematically review the literature on animal host species and geographic distribution of Leptospira serovars to examine associations between serovars with animal host species and regions, and to identify geographic regions in need of study. Methods: Nine library databases were searched from inception through 9 March 2023 using keywords including Leptospira, animal, and a list of serovars. We sought reports of detection of Leptospira, from any animal, characterized by cross agglutinin absorption test, monoclonal antibody typing, serum factor analysis, or pulsed-field gel electrophoresis to identify the serovar. Results: We included 409 reports, published from 1927 through 2022, yielding data on 154 Leptospira serovars. The reports included data from 66 (26.5%) of 249 countries. Detections were from 144 animal host species including 135 (93.8%) from the class Mammalia, 5 (3.5%) from Amphibia, 3 (2.1%) from Reptilia, and 1 (0.7%) from Arachnida. Across the animal host species, Leptospira serovars that were detected in the largest number of animal species included Grippotyphosa (n=39), Icterohaemorrhagiae (n=29), Pomona (n=28), Australis (n=25), and Ballum (n=25). Of serovars, 76 were detected in a single animal host species. We created an online database to identify animal host species for each serovar by country. Conclusions: We found that many countries have few or no Leptospira serovars detected from animal host species and that many serovars were detected from a single animal species. Our study highlights the importance of efforts to identify animal host species of leptospirosis, especially in places with a high incidence of human leptospirosis. We provide an updated resource for leptospirosis researchers.
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Rapid and simple serologic tests that require only a small amount of blood without the euthanization of animals are valuable for microbial control in colonies of laboratory animals. In this study, we developed a multiplex immunochromatographic assay (ICA) for detection of antibodies to Sendai virus (also known as hemagglutinating virus of Japan), hantavirus, and sialodacryoadenitis virus, which are causative agents of major infectious diseases in rats. For this assay, an ICA strip was placed into a microtube containing 150 µl PBS and either 0.75 µl of rat serum or 1.5 µl of whole blood. Binding antibodies were visualized by using anti-rat IgG antibody-conjugated colloidal gold. Under these conditions, the multiplex ICA simultaneously and specifically detected antibodies to multiple antigens. Positive serum samples for each infectious disease were used to evaluate the sensitivity and specificity of the multiplex ICA. The sensitivities of the multiplex ICA for Sendai virus, hantavirus, and sialodacryoadenitis virus were 100%, 100%, and 81%, respectively. No nonspecific reactions were observed in any of the 52 positive sera against heterologous antigens. In addition, 10 samples of uninfected sera did not show any bands except for the control line. These observations indicate high specificity of the multiplex ICA. Moreover, the multiplex ICA could be applied to diluted blood. These results indicate that the multiplex ICA is appropriate for rapid and simple serological testing of laboratory rats.
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Infecções por Coronavirus/diagnóstico , Coronavirus do Rato/isolamento & purificação , Infecções por Hantavirus/diagnóstico , Imunoensaio/métodos , Orthohantavírus/isolamento & purificação , Infecções por Respirovirus/diagnóstico , Doenças dos Roedores/diagnóstico , Vírus Sendai/isolamento & purificação , Animais , Ratos , Testes SorológicosRESUMO
In susceptible hosts, protection from Leptospira infection is mediated by the innate immune response at the point of entry and humoral immunity. Thus, identifying and segregating the initial host response at the representative host-pathogen interface is needed to understand the typical outcomes of Leptospira infection, clearance, persistence, or disease. An in vitro whole blood culture system to study the overall immune response using pathogenic and non-pathogenic Leptospira strains was explored in this study. Using an ELISA, increased IL-8, TNF alpha, and IL-1 in blood samples stimulated with pathogenic and nonpathogenic Leptospira compared to unstimulated controls were detected. In RT2 Profiler PCR Array assays, consistent upregulation of 22 genes and downregulation of 25 genes were observed. Few of the notable upregulated genes included BPI, CCL3, CXCL2, IL-6, IL-8, TLR1, TLR2, TLR6, and TNF and downregulated genes included, LBP, LYZ, MPO, MYD88. IFNß was upregulated in samples treated with pathogenic Leptospira and IL-1ß was upregulated in samples treated with nonpathogenic Leptospira. Toll- like Receptor signaling and expression of pattern recognition receptors were two of the five prominent canonical pathways observed. Individual deconvolution of each of the specific and significant pathways observed in this study may improve the understanding of the pathogenesis of this important zoonotic agent. The use of this system in conjunction with whole transcriptome analysis in a larger population, may unveil the robust nature of host/Leptospira interaction.
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Hemocultura/veterinária , Doenças do Cão/imunologia , Leptospira/imunologia , Leptospirose/veterinária , Animais , Hemocultura/métodos , Doenças do Cão/microbiologia , Cães , Imunidade Inata , Leptospirose/imunologia , Leptospirose/microbiologia , Receptores Toll-Like/genéticaRESUMO
This study was performed to investigate the potential asymptomatic Leptospira reservoir status among African green monkeys (AGMs) in the Caribbean island of Saint Kitts, and whether there is any renal pathology associated with Leptospira exposure. Forty-eight percent of AGMs tested were positive for Leptospira antibodies by the microscopic agglutination test. Leptospira DNA was detected in 4% of kidney samples tested using a lipl32 gene based PCR. We observed minimal to severe microscopic renal lesions in 85% of the AGM kidneys evaluated. The majority of the AGMs (n = 26) had only minimal to mild interstitial nephritis and a few (n = 3) had moderate to severe lesions. The presence of interstitial nephritis was not significantly associated with Leptospira exposure. The presence of infected AGMs in a small surface limited geographic region may pose zoonotic threat to humans and animals. The impact of Leptospira infection in renal pathology in AGMs warrants further investigation. AGMs residing in a natural setting in an insular, surface limited Leptospira endemic geographic region may offer opportunities for comparative studies to advance the field of leptospirosis. Due to their similarity to humans, such studies in AGMs may also provide translational opportunities to advance Leptospira research.
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In this study, we detected and characterized Leptospira infection and exposure in rats on the Caribbean island of Saint Kitts for the first time. We detected Leptospira infection in 17/29 (59%), 14/29 (48)%, and 11/29 (38)% of rats by RT-PCR, culture, and immunofluorescence assay, respectively. Whole genome sequencing (WGS) and analysis and serogrouping of 17 Leptospira strains isolated from rats revealed their close relationship with L. interrogans serogroup Icterohaemorrhagiae (n = 10) and L. borgpetersenii serogroup Ballum (n = 7). WGS, serogrouping, and additional PCR tests on rat kidneys confirmed mixed infections with L. interrogans and L. borgpetersenii in the kidneys of three rats. Microscopic agglutination test (MAT) was positive for 25/29 (87%) of the rats tested, and the response was restricted to serovars Icterohaemorrhagiae {24/29(83%)}, Mankarso {4/29(14%)}, Copenhageni {4/29(14%)}, Grippotyphosa {2/29(7%)}, and Wolffi {1/29(3%)}. Interestingly, there was no agglutinating antibody response to serovar Ballum. We observed a similar pattern in the serologic response using Leptospira isolates obtained from this study with each of the rat sera, with strong response to L. interrogans isolates but minimal reactivity to L. borgpetersenii isolates. Our findings suggest the use of multiple complementary diagnostic tests for Leptospira surveillance and diagnosis to improve the accuracy of the data.
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Serologic monitoring of infectious diseases is important for microbial control in colonies of laboratory mice. Rapid and simple tests that do not require killing animals are valuable for this purpose. In this study, we developed a multiplex immunochromatographic assay (ICA) for detection of antibodies to mouse hepatitis virus (MHV), Sendai virus (also known as hemagglutinating virus of Japan [HVJ]), and Clostridium piliforme (The pathogen that causes Tyzzer disease), which are major infectious diseases in mice. For this assay, an ICA strip was put into a microtube containing 150 µL PBS and either 0.75 µL mouse serum or 1.5 µL whole blood. Binding antibodies were visualized by using protein A-conjugated colloidal gold. Under these conditions, multiplex ICA simultaneously and specifically detected antibodies to multiple antigens. To evaluate the sensitivity and specificity of multiplex ICA, positive serum samples for each infectious disease were used. Sensitivities of the multiplex ICA test for MHV, HVJ, and C. piliforme were 100%, 100%, and 90%, respectively. No nonspecific reaction was observed in any of the 30 positive sera. In addition, 10 samples of uninfected sera did not show any bands except for the control line. These observations indicate high specificity of the multiplex ICA test. Moreover, the multiplex ICA could be applied to diluted blood. These results indicate that the multiplex ICA is appropriate for rapid, simple, and safe serologic testing of laboratory mice.
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Doenças Transmissíveis/veterinária , Imunoensaio/veterinária , Doenças dos Roedores/diagnóstico , Animais , Animais de Laboratório , Doenças Transmissíveis/sangue , Doenças Transmissíveis/diagnóstico , Imunoensaio/métodos , Ciência dos Animais de Laboratório , Camundongos , Doenças dos Roedores/sangue , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: The role of rodents in Leptospira epidemiology and transmission is well known worldwide. Rats are known to carry different pathogenic serovars of Leptospira spp. capable of causing disease in humans and animals. Wild rats (Rattus spp.), especially the Norway/brown rat (Rattus norvegicus) and the black rat (R. rattus), are the most important sources of Leptospira infection, as they are abundant in urban and peridomestic environments. In this study, we compiled and summarized available data in the literature on global prevalence of Leptospira exposure and infection in rats, as well as compared the global distribution of Leptospira spp. in rats with respect to prevalence, geographic location, method of detection, diversity of serogroups/serovars, and species of rat. METHODS: We conducted a thorough literature search using PubMed without restrictions on publication date as well as Google Scholar to manually search for other relevant articles. Abstracts were included if they described data pertaining to Leptospira spp. in rats (Rattus spp.) from any geographic region around the world, including reviews. The data extracted from the articles selected included the author(s), year of publication, geographic location, method(s) of detection used, species of rat(s), sample size, prevalence of Leptospira spp. (overall and within each rat species), and information on species, serogroups, and/or serovars of Leptospira spp. detected. FINDINGS: A thorough search on PubMed retrieved 303 titles. After screening the articles for duplicates and inclusion/exclusion criteria, as well as manual inclusion of relevant articles, 145 articles were included in this review. Leptospira prevalence in rats varied considerably based on geographic location, with some reporting zero prevalence in countries such as Madagascar, Tanzania, and the Faroe Islands, and others reporting as high as >80% prevalence in studies done in Brazil, India, and the Philippines. The top five countries that were reported based on number of articles include India (n = 13), Malaysia (n = 9), Brazil (n = 8), Thailand (n = 7), and France (n = 6). Methods of detecting or isolating Leptospira spp. also varied among studies. Studies among different Rattus species reported a higher Leptospira prevalence in R. norvegicus. The serovar Icterohaemorrhagiae was the most prevalent serovar reported in Rattus spp. worldwide. Additionally, this literature review provided evidence for Leptospira infection in laboratory rodent colonies within controlled environments, implicating the zoonotic potential to laboratory animal caretakers. CONCLUSIONS: Reports on global distribution of Leptospira infection in rats varies widely, with considerably high prevalence reported in many countries. This literature review emphasizes the need for enhanced surveillance programs using standardized methods for assessing Leptospira exposure or infection in rats. This review also demonstrated several weaknesses to the current methods of reporting the prevalence of Leptospira spp. in rats worldwide. As such, this necessitates a call for standardized protocols for the testing and reporting of such studies, especially pertaining to the diagnostic methods used. A deeper understanding of the ecology and epidemiology of Leptospira spp. in rats in urban environments is warranted. It is also pertinent for rat control programs to be proposed in conjunction with increased efforts for public awareness and education regarding leptospirosis transmission and prevention.
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Leptospirose/epidemiologia , Leptospirose/veterinária , Ratos/microbiologia , Animais , Bases de Dados Factuais , Mapeamento Geográfico , Leptospira/classificação , Leptospira/genética , Leptospira/isolamento & purificação , Leptospirose/diagnóstico , Prevalência , SorogrupoRESUMO
A pilot seroprevalence study was conducted to document exposure to selected pathogens in wild rats inhabiting the Caribbean island of St. Kitts. Serum samples collected from 22 captured wild rats (Rattus norvegicus and Rattus rattus) were tested for the presence of antibodies to various rodent pathogens using a rat MFI2 serology panel. The samples were positive for cilia-associated respiratory bacillus (13/22; 59.1%), Clostridium piliforme (4/22; 18.2%), Mycoplasma pulmonis (4/22; 18.2%), Pneumocystis carinii (1/22; 4.5%), mouse adenovirus type 2 (16/22; 72.7%), Kilham rat virus (15/22; 68.2%), reovirus type 3 (9/22; 40.9%), rat parvovirus (4/22; 18.2%), rat minute virus (4/22; 18.2%), rat theilovirus (2/22; 9.1%), and infectious diarrhea of infant rats strain of group B rotavirus (rat rotavirus) (1/22; 4.5%). This study provides the first evidence of exposure to various rodent pathogens in wild rats on the island of St. Kitts. Periodic pathogen surveillance in the wild rat population would be beneficial in assessing potential regional zoonotic risks as well as in enhancing the current knowledge when implementing routine animal health monitoring protocols in facilities with laboratory rodent colonies.
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This pilot study describes the prevalence of Leptospira infection and exposure in livestock species, cattle, pig, sheep, and goats in Saint Kitts in the Caribbean region. Serum and kidney samples were collected from cattle, pigs, sheep, and goats at a local abattoir between September 2016 and March 2017. Cattle had the highest seroprevalence (79.8%) followed by pigs (64.8%), sheep (39.4%), and goats (24.8%). Highest seroprevalence was observed to serovars, Mankarso in cattle, Bratislava in pigs, Hardjo in sheep, and goats. Leptospira DNA was amplified from kidney samples of 18/99 cattle (18.2%), 11/106 pigs (10.4%), 4/106 sheep (3.8%), and 2/105 goats (1.9%). Our findings warrant further studies to assess leptospirosis associated economic burden to subsistence farmers and public health impact.
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Leptospira/isolamento & purificação , Leptospirose/veterinária , Gado , Animais , Leptospira/genética , Leptospirose/epidemiologia , Leptospirose/microbiologia , Projetos Piloto , Prevalência , São Cristóvão e Névis/epidemiologia , Estudos Soroepidemiológicos , SorogrupoRESUMO
In this study, we investigated Leptospira infection and exposure in small Indian mongoose (Herpestes auropunctatus), an invasive animal species, in two different sites in the Caribbean island of Saint Kitts. Overall a low seroprevalence (12/148; 8.1%: 95%CI: 3.7-12.5) was observed. Agglutinating antibodies to serovars Mankarso (3.4%), Copenhageni (2.7%), Icterohemorrhagiae (1.4%), Bratislava (1.4%), Canicola (1.4%), Autumnalis (0.7%), Alexi (0.7%), Pomona (0.7%) and Grippotyphosa (0.7%) was observed on the microscopic agglutination test. The seroprevalence observed in mongooses trapped from peridomestic sites was significantly higher compared to the arid and less inhabited site (p = 0.0268). The real time PCR targeting lipL32 gene was positive for 9 out of 146 mongooses. Bacterial culture of kidneys resulted in two Leptospira isolates. Whole genome sequencing and analysis suggested that these isolates are closely related to L. interrogans serovar Copenhageni. We observed mild to severe chronic renal lesions in 20.2% of mongooses in the absence of an antibody response or active infection. Our findings emphasize the need to investigate other infectious etiologies or atypical outcomes and potential chronic long-term impact of Leptospira infection in animals and people living in an endemic area. In addition, our data reinforces the need for including locally prevalent Leptospira isolates rather than representative members of a serogroup in the microscopic agglutination test panel in epidemiologic and diagnostic investigations. In conclusion, mongoose inhabiting the island are exposed to and harbor pathogenic Leptospira and hence may play a role in the transmission. The invasive nature of the species highlights their presence as a potential risk factor for this widespread zoonotic disease.
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Herpestidae , Nefropatias/microbiologia , Leptospira interrogans/genética , Leptospirose/epidemiologia , Testes de Aglutinação , Animais , Anticorpos Antibacterianos/sangue , Espécies Introduzidas , Nefropatias/patologia , Leptospira interrogans/isolamento & purificação , Leptospirose/diagnóstico , Prevalência , Fatores de Risco , Estudos Soroepidemiológicos , Sorogrupo , Índias Ocidentais/epidemiologia , Zoonoses/epidemiologiaRESUMO
Monoclonal antibody (MAb)-based ELISA was developed to detect leptospiral antigens from the plasma of febrile patients. The MAb reacts specifically with pathogenic leptospires and the assay possesses excellent diagnostic test parameters compared to PCR, indicating that this new ELISA is useful for the early diagnosis of leptospirosis with low operational cost.
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Anticorpos Antibacterianos/química , Anticorpos Monoclonais Murinos/química , Antígenos de Bactérias/imunologia , Leptospira interrogans/imunologia , Leptospirose , Ensaio de Imunoadsorção Enzimática/métodos , Humanos , Leptospirose/diagnóstico , Leptospirose/imunologia , Sensibilidade e EspecificidadeRESUMO
We report here high rates of detection (54%, 21/39) of picobirnaviruses (PBVs) in feces/intestinal contents of free roaming, apparently healthy rats (Rattus spp.) on the Caribbean island of St. Kitts. One of the PBV strains, strain PBV/Rat/KNA/Rat9/2017, was molecularly characterized for complete gene segment-2. To determine the nucleotide (nt) sequence of full-length gene segment-2, the 5'- and 3'- portions of gene segment-2 of strain Rat9 containing an overlapping region were amplified using a non-specific primer-based amplification method with modifications. The complete gene segment-2 of PBV strain Rat9 was 1730â¯bp in length, encoding a putative RNA-dependent RNA polymerase (RdRp) of 535 amino acid (aa). By nt and deduced aa sequence identities and phylogenetic analysis, the complete gene segment-2 of strain Rat9 exhibited high genetic diversity with those of PBVs from other host species. On the other hand, 5'- and 3'- end nt sequences of gene segment-2, and the three domains of putative RdRp that are conserved in PBVs were retained in strain Rat9. To our knowledge, this is the first report on molecular characterization of complete gene segment-2 of a PBV strain from Rattus spp., providing important insights into the putative RdRp, and genetic diversity and evolution of PBV in rats. The high detection rates of PBV in free roaming rats on St. Kitts emphasizes the importance of further studies on epidemiology and genetic makeup of PBVs in Rattus spp.
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Doenças dos Animais/virologia , Picobirnavirus/genética , Infecções por Vírus de RNA/veterinária , Sequência de Aminoácidos , Animais , Genes Virais , Genoma Viral , Filogenia , Picobirnavirus/classificação , RNA Polimerase Dependente de RNA/química , RNA Polimerase Dependente de RNA/genética , RatosRESUMO
We examined 33 rodents captured in an urban area of Osaka City, Japan for IgG antibodies against Seoul virus, severe fever with thrombocytopenia syndrome virus, hepatitis E virus, Leptospira interrogans, Yersinia pestis, spotted fever, typhus and scrub typhus group rickettsiae. We found that 3 (9.1%) and 1 (3.0%) of the 33 rodents had antibodies against L. interrogans and spotted fever group rickettsiae, respectively. DNAs of leptospires were detected from 2 of the 3 seropositive rodents, but DNA of rickettsia was not detected. Phylogenetic analysis and multiple locus sequence typing revealed that the 2 leptospires were L. interrogans belonging to a novel sequence type. There is a potential risk for acquiring rodent-borne zoonotic pathogens even in cities in developed countries.
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Leptospira interrogans , Leptospirose/veterinária , Ratos/microbiologia , Rickettsia , Rickettsiose do Grupo da Febre Maculosa/veterinária , Animais , Cidades , DNA Bacteriano/genética , Japão/epidemiologia , Leptospira interrogans/genética , Leptospirose/epidemiologia , Leptospirose/microbiologia , Tipagem de Sequências Multilocus/veterinária , Filogenia , Rickettsia/genética , Rickettsiose do Grupo da Febre Maculosa/epidemiologia , Rickettsiose do Grupo da Febre Maculosa/microbiologiaRESUMO
The applicability of the recombinant LipL32 for serodiagnosis of leptospiral infection in field rodents was assessed in this study. An immunodominant region of LipL32 was determined by monoclonal antibodies, and then, truncated LipL32 (tLipL32) was designed to contain the region (87-188th amino acid). The tLipL32 was compared between two recombinant expression hosts Escherichia coli and Pichia pastoris in ELISA. With field rat sera, tLipL32 expressed by P. pastoris (tLipL32p) had high antigenicity without background reactions, while tLipL32 expressed by E. coli (tLipL32e) showed high background reactions, which were reduced by pre-adsorption of sera with E. coli. To evaluate tLipL32-ELISA, field rat sera were tentatively divided into a Leptospira infection positive (12 sera) and a negative group (12 sera) based on the results from flaB gene PCR of kidney samples and WB with whole Leptospira cell. Consequently, the sensitivity of tLipL32p-ELISA for field rat sera was 83% . A similar result was obtained from tLipL32e-ELISA with adsorbed sera, (92%). However, sensitivity of tLipL32e-ELISA using sera without an adsorption treatment was 50%. Regardless of the expression host, tLipL32-ELISA had 100% specificity and sensitivity in experimentally infected laboratory rats. These results suggest that recombinant LipL32 expressed by P. pastoris is more applicable for serodiagnosis in field rats due to a lack of background reaction.
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Proteínas da Membrana Bacteriana Externa , Leptospirose/diagnóstico , Lipoproteínas , Animais , Anticorpos Antibacterianos/imunologia , Anticorpos Monoclonais/imunologia , Proteínas da Membrana Bacteriana Externa/genética , Proteínas da Membrana Bacteriana Externa/imunologia , Ensaio de Imunoadsorção Enzimática , Mapeamento de Epitopos , Escherichia coli/genética , Feminino , Leptospira/imunologia , Lipoproteínas/genética , Lipoproteínas/imunologia , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/imunologia , Pichia/genética , Ratos , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Testes SorológicosRESUMO
BACKGROUND: Hantaviruses are causative agents of hemorrhagic fever with renal syndrome (HFRS) and nephropathia epidemica (NE) in the Old World and hantavirus pulmonary syndrome (HPS) in the New World. There is a need for time-saving diagnostic methods. In the present study, recombinant N antigens were used as antigens in an immunochromatography strip (ICG) test to detect specific IgG antibodies. METHODS: The N-terminal 103 amino acids (aa) of Hantaan virus (HTNV), Puumala virus (PUUV) and Andes virus (ANDV) nucleocapsid (N) protein were expressed in E. coli as representative antigens of three groups (HFRS, NE and HPS-causing viruses) of hantavirus. Five different types of ICG test strips, one antigen line on one strip for each of the three selected hantaviruses (HTNV, PUUV and ANDV), three antigen lines on one strip and a mixed antigen line on one strip, were developed and sensitivities were compared. RESULTS: A total of 87 convalescent-phase patient sera, including sera from 35 HFRS patients, 36 NE patients and 16 HPS patients, and 25 sera from healthy seronegative people as negative controls were used to evaluate the ICG test. Sensitivities of the three-line strip and mixed-line strip were similar to those of the single antigen strip (97.2 to 100%). On the other hand, all of the ICG test strips showed high specificities to healthy donors. CONCLUSION: These results indicated that the ICG test with the three representative antigens is an effective serodiagnostic tool for screening and typing of hantavirus infection in humans.