Cynoglossum officinale, a New Natural Host of Alfalfa mosaic virus

Cynoglossum officinale L. (family Boraginaceae), hound's-tongue, is a medicinal plant whose roots are used for their astringent and healing properties and for their sedative, calming, and slightly narcotic effects. This species, originating from Europe where it grows wild in mountainous fields, is cultivated only for its medicinal properties. Epidemiological surveys performed in the Emilia Romagna Region of northern Italy in the spring and autumn of 2001 revealed the presence of virus-like symptoms on C. officinale cultivated in two different locations. In the Botanical Garden of the University of Bologna, the plants showed stunting, interveinal chlorotic spots, midrib necrosis, and scarce or no seed production. In the experimental field of the Agriculture Faculty of Bologna (Imola), the plants exhibited stunting, interveinal chlorotic spots, and reduction of leaf lamina. Mechanical inoculations of sap from symptomatic leaves caused typical symptoms of Alfalfa mosaic virus (AlMV) on Chenopodium amaranticolor Coste & Reyn. (local chlorotic, necrotic lesions and systemic chlorotic vein streaking), Vigna unguicolata (L.) Walp., and Phaseolus vulgaris L. (local necrotic lesions). Using an electron microscope, examination of leaf sap obtained from infected plants stained with uranyl acetate and phosphotungstic acid did not show the presence of elongated virus particles. Serological tests, such as immunoelectron microscopy, gold-labeled decoration, and protein A sandwich indirect enzyme-linked immunosorbent assay using antiserum to AMV (PVAS 92, American Type Culture Collection, Manassas, VA, and AlMV-Vinca minor L., from the DiSTA collection, Bologna, Italy as a control), gave positive reactions, indicating that the virus in question was AlMV. To our knowledge, this is the first report of virus infection on C. officinale, a new natural host of AlMV.

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First Record of Alfalfa mosaic virus in Teucrium fruticans in Italy

Plant Disease ◽

10.1094/pdis-08-11-0674 ◽

2012 ◽

Vol 96 (2) ◽

pp. 294-294 ◽

Cited By ~ 2

Author(s):

G. Parrella ◽

L. Cavicchi ◽

M. G. Bellardi

Keyword(s):

Mosaic Virus ◽

Consensus Sequence ◽

Protein A ◽

Alfalfa Mosaic Virus ◽

Pairwise Comparison ◽

Geographic Area ◽

Rt Pcr ◽

Leaf Extracts ◽

Chenopodium Amaranticolor ◽

Bright Yellow

The Teucrium genus (Lamiaceae family) contains ~300 species of evergreen and deciduous shrubs with some species widely used as ornamental plants in rock gardens. During the springs of 2010 and 2011, some plants of Teucrium fruticans L., also known as “tree germander”, growing singly in pots in a Ligurian nursery (Savona Province, northern Italy), were noted for a bright yellow calico mosaic on the leaves (~1% of ~2,000 plants inspected exhibited symptoms). Preliminary electron microscope observations of leaf-dips showed semispherical to bacilliform particles, consistent with Alfamovirus and Oleavirus, in preparations obtained only from leaves of symptomatic plants. Three symptomatic and two asymptomatic plants were checked for Cucumber mosaic virus or Alfalfa mosaic virus (AMV) in protein A sandwich (PAS)-ELISA with commercial kits (Bioreba, Reinach, Switzerland) and Olive latent virus 2 (OLV2) by immunodecoration of virus particles with an OLV2 antiserum produced against an Italian OLV2 isolate. Symptomatic plants were positive only to AMV and all asymptomatic plants were negative to all viruses checked. The virus was successfully transmitted mechanically to Chenopodium amaranticolor and Ocimum basilicum that reacted, as expected for infections caused by AMV (1), with a chlorotic local lesion followed by mosaic and bright yellow mosaic, respectively. The disease was transmitted also by grafting an infected scion on healthy T. fruticans. Symptoms appeared after ~3 weeks in one plant of six grafted. AMV infection in a symptomatic grafted plant was verified by PAS-ELISA, confirming that bright yellow mosaic symptoms observed in T. fruticans were induced by an isolate of AMV. Immunocapture reverse transcription (IC-RT)-PCR assay, following the protocol described by Wetzel et al. (4), was performed on leaf extracts from one symptomatic plant using a polyclonal serum raised against a French isolate of AMV, provided by H. Lot (INRA, Station de Pathologie Végétale, Avignon, France). Specific AMV primer pair was used in the RT-PCR reactions (2). A DNA fragment of ~750 bp, covering the entire coat protein gene (CP), was obtained after IC-RT-PCR. The amplicon was gel purified with the Wizard SV Gel and PCR Clean-Up System (Promega, Madison, WI), cloned into pGEMT-easy vector (Promega) and two independent clones sequenced on both strands at MWG Biotech (Ebersberg, Germany). The consensus sequence was submitted to EMBL (No. FR854391). Pairwise comparison of the AMV-T. fruticans isolate CP sequence (named Tef-1) with those of AMV reference isolates revealed the maximum (98.0 to 97.3%) nucleotide identities with isolates belonging to subgroup I, 95.5 to 94.0% identities with subgroup IIA isolates, and 95.6% identity with the subgroup IIB isolate Tec-1 (3). Among subgroup I isolates, Tef-1 had the maximum CP nucleotide identity with the CP gene belonging to an AMV isolate identified in 2010 in Lavandula stoechas in the same geographic area, suggesting a common origin for these two viral isolates. Overall results clearly indicate that an AMV isolate was the causal agent of the calico-type mosaic observed in T. fruticans. To our knowledge, this is the first report of T. fruticans as a natural host of AMV. References: (1) G. Marchoux et al. Page 163 in: Virus des Solanacées. Quae éditions, Versailles, 2008. (2) G. +Parrella et al. Arch. Virol. 145:2659, 2000. (3) G. Parrella et al. Arch. Virol. 156:1049, 2011. (4) T. Wetzel et al. J. Virol. Methods 39:27, 1992.

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Identification, Characterization, and Molecular Detection of Alfalfa mosaic virus in Potato

Phytopathology ◽

10.1094/phyto-96-1237 ◽

2006 ◽

Vol 96 (11) ◽

pp. 1237-1242 ◽

Cited By ~ 34

Author(s):

H. Xu ◽

J. Nie

Keyword(s):

Mosaic Virus ◽

Gene Sequence ◽

Alfalfa Mosaic Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Polymorphism Analysis ◽

Rt Pcr ◽

Potato Leaf ◽

Simple Method ◽

Cp Gene ◽

Nucleotide Sequence Alignment

Alfalfa mosaic virus (AMV) was detected in potato fields in several provinces in Canada and characterized by bioassay, enzyme-linked immunosorbent assay, and reverse-transcription polymerase chain reaction (RT-PCR). The identity of eight Canadian potato AMV isolates was confirmed by sequence analysis of their coat protein (CP) gene. Sequence and phylogenetic analysis indicated that these eight AMV potato isolates fell into one strain group, whereas a slight difference between Ca175 and the other Canadian AMV isolates was revealed. The Canadian AMV isolates, except Ca175, clustered together among other strains based on alignment of the CP gene sequence. To detect the virus, a pair of primers, AMV-F and AMV-R, specific to the AMV CP gene, was designed based on the nucleotide sequence alignment of known AMV strains. Evaluations showed that RT-PCR using this primer set was specific and sensitive for detecting AMV in potato leaf and tuber samples. AMV RNAs were easily detected in composite samples of 400 to 800 potato leaves or 200 to 400 tubers. Restriction analysis of PCR amplicons with SacI was a simple method for the confirmation of PCR tests. Thus, RT-PCR followed by restriction fragment length polymorphism analysis may be a useful approach for screening potato samples on a large scale for the presence of AMV.

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First Report of Soilborne Wheat Mosaic Virus in the United Kingdom

Plant Disease ◽

10.1094/pdis.1999.83.9.880a ◽

1999 ◽

Vol 83 (9) ◽

pp. 880-880 ◽

Cited By ~ 9

Author(s):

G. R. G. Clover ◽

D. M. Wright ◽

C. M. Henry

Keyword(s):

United Kingdom ◽

Mosaic Virus ◽

Barley Yellow Dwarf Virus ◽

Protein A ◽

The United States ◽

Enzyme Linked Immunosorbent Assay ◽

Barley Yellow Dwarf ◽

The United Kingdom ◽

Field Samples ◽

Yellow Dwarf Virus

In April 1999, severe soilborne wheat mosaic virus (SBWMV) symptoms were observed in five fields of winter wheat (Triticum aestivum, cvs. Consort, Equinox, and Savannah) on one farm in Wiltshire, UK. Affected plants were markedly stunted and had a pale mosaic on their leaf sheaths that developed into bright yellow, parallel streaks on the leaves as they unfolded. Symptomatic plants were found in discrete, elliptical patches ranging in size from a few square meters to nearly a hectare. During May and June, symptoms became less marked as temperatures increased and were restricted to lower leaves. SBWMV was positively identified in all five fields (60 to 170 plants per field) by double (W. Huth, BBA-Braunschweig, Germany; Sanofi Phyto-Diagnostics, Paris) and triple (T. Wilson, SCRI, Dundee, UK) antibody sandwich enzyme-linked immunosorbent assay and by reversetranscription polymerase chain reaction (2). Identification was confirmed by immunoelectron microscopy, including protein-A gold labeling, which revealed bipartite, rod-shaped particles typical of SBWMV. Neither wheat spindle streak mosaic virus nor barley yellow dwarf virus was detected in the field samples, nor was SBWMV detected in any other field subsequently sampled, despite a survey of the surrounding area. Wheat is the most important economic crop in the United Kingdom (≈1.9 million hectares are grown annually, yielding ≈16 million tonnes), but its position is threatened by the economic impact of SBWMV, which has decreased yields by up to 50% in the United States (1). References: (1) T. A. Kucharek and J. H. Walker. Plant Dis. Rep. 58:763, 1974. (2) R. E. Pennington et al. Plant Dis. 77:1202, 1993.

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First Report of Pepino mosaic virus on Tomato in Spain

Plant Disease ◽

10.1094/pdis.2001.85.12.1292c ◽

2001 ◽

Vol 85 (12) ◽

pp. 1292-1292 ◽

Cited By ~ 5

Author(s):

C. Jordá ◽

A. Lázaro Pérez ◽

P. Martínez-Culebras ◽

P. Abad ◽

A. Lacasa ◽

...

Keyword(s):

Mosaic Virus ◽

Canary Islands ◽

Plant Protection ◽

Polyclonal Antibodies ◽

Datura Stramonium ◽

Enzyme Linked Immunosorbent Assay ◽

Chenopodium Amaranticolor ◽

Pepino Mosaic Virus ◽

Tomato Plants ◽

First Report

At the beginning of 2000, a damaging disease developed on protected tomato (Lycopersicon esculentum) crops grown in polyethylene greenhouses in different regions of Spain. Production losses were estimated at 15 to 80%. The tomato plants showed a variety of symptoms. The most common symptoms were leaf distortion, chlorosis, and mosaic. Some plants showed a dark green mosaic and bubbling of the leaf surface. Green striations were also observed on the stem and sepals. Most of the diseased plants had discolored fruits. Symptoms decreased as environmental temperature increased. The involvement of Pepino mosaic virus (PepMV) was suspected. To identify the etiological agent, ≈500 symptomatic tomato plants were collected from several locations in Alicante, Murcia, Almeria and the Canary Islands. Flexuous viral particles 510 nm long were observed by transmission electron microscopy, suggesting the presence of a potexvirus in the tissue extracts analyzed. All samples were tested by ELISA (enzyme-linked immunosorbent assay), using polyclonal antibodies to Narcissus mosaic virus (Adgen, Auchincriuve, Scotland), a virus serologically related to PepMV, and two antisera specific to PepMV (Adgen, Scotland and DMSZ, Braunschweig, Germany). PepMV was detected in 35% of the samples. Like PepMV, the virus infected (as confirmed by ELISA) greenhouse-grown Datura stramonium, Nicandra physalodes, Nicotiana benthamiana, N. clevelandii, Solanum tuberosum, and Vigna sinensis and did not infect Capsicum anuum, Cucumis sativus, Chenopodium amaranticolor, C. quinoa, Petunia × hybrida, Phaseolus vulgaris, Physalis floridana, N. glutinosa, N. rustica, or N. tabacum. The virus did infect Gomphrena globosa, which normally is not infected by PepMV. The first report of PepMV was on pepino (Solanum muricatum) in Peru in 1974 (1), but this virus has been recently reported in the Netherlands, England, Germany, and France on protected tomato crops (2). To our knowledge, this is the first report of PepMV in Spain, including the Canary Islands. References: (1) R. A. C. Jones et al. Ann. Appl. Biol. 94:61, 1980. (2) European and Mediterranean Plant Protection Organisation (EPPO). Alert List Viruses. On-line publication/2000/003.

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First Report of Alfalfa mosaic virus in Lavandula officinalis

Plant Disease ◽

10.1094/pdis.2004.88.8.908c ◽

2004 ◽

Vol 88 (8) ◽

pp. 908-908 ◽

Cited By ~ 6

Author(s):

Ll. Martínez-Priego ◽

M. C. Córdoba ◽

C. Jordá

Keyword(s):

Mosaic Virus ◽

Screening Program ◽

Sequence Data ◽

Alfalfa Mosaic Virus ◽

Mediterranean Area ◽

Enzyme Linked Immunosorbent Assay ◽

First Report ◽

Coat Protein Region ◽

C 30 ◽

Lavandula Officinalis

For several years, in ornamental nurseries in the Mediterranean area of Spain, stunting and yellow leaf spotting have been observed in young plants of Lavandula officinalis. Symptoms eventually disappeared as the plants matured. During the summer of 2003, the number of plantlets affected and the intensity of symptoms increased significantly. Symptomatic plants tested positive using enzyme-linked immunosorbent assay (ELISA) (Phyto-Diagnostics, INRA, France) for the presence of Alfalfa mosaic virus (AMV). ELISA results were verified using reverse transcription-polymerase chain reaction (RT-PCR). Total RNA extracts from symptomatic plants were analyzed using primers designed specifically for the coat protein region of AMV utilizing sequence data from GenBank Accession No. AF215664: AMVcoat-F: GT GGT GGG AAA GCT GGT AAA and AMVcoat-R: CAC CCA GTG GAG GTC AGC ATT. The thermocycling schedule was as follows: reverse transcriptase step at 50°C for 30 min, first PCR cycle at 94°C for 2 min, 35 cycles each of 30 s at 94°C, 30 s at 54°C, 30 s at 72°C, followed by a final extension at 72°C for 10 min. A 700-pb PCR product of the expected size was obtained from plants that were positive for AMV using ELISA. The two systems provide for rapid detection of AMV in L. officinalis. A regular screening program will assist in providing virus-free plants to ornamental nurseries. These results demonstrate the presence of AMV in L. officinalis. Alfalfa (Medicago sativa L.) is a typical source of AMV. However, because the nurseries where L. officinalis is grown are not in the vicinity of alfalfa fields, we suggest the source of the infection originated in the propagation material. AMV has currently been reported in L. officinalis only in Italy and France (1). To our knowledge, this is the first report of AMV in L. officinalis in Spain. Reference: (1): A. Garibaldi et al. Ed. Edagricole-Edisioni Agricole della Calderini s.r.l., Bologna, 2000.

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A Multiyear Survey and Identification of Pepper- and Tomato-Infecting Viruses in Yunnan Province, China

Frontiers in Microbiology ◽

10.3389/fmicb.2021.623875 ◽

2021 ◽

Vol 12 ◽

Author(s):

Yueyue Li ◽

Guanlin Tan ◽

Long Xiao ◽

Wenpeng Zhou ◽

Pingxiu Lan ◽

...

Keyword(s):

Mosaic Virus ◽

Mixed Infection ◽

Yunnan Province ◽

Virus Disease ◽

Natural Host ◽

Tomato Mosaic Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Tomato Production ◽

Infection Types ◽

First Time

During pepper and tomato production seasons in 2013–2017, large-scale virus disease surveys were conducted in different regions of Yunnan Province, China. A total of 1,267 pepper and tomato samples with various virus-like symptoms were collected and analyzed for virus infections through dot enzyme-linked immunosorbent assay (dot-ELISA), polymerase chain reaction (PCR), and reverse-transcription (RT)-PCR. The detection results showed that 19 different viruses were present in about 50.9% of the assayed samples, and among these viruses, seven viruses were found in both pepper and tomato samples. Mixed infections with two to three of the 15 identified mixed infection types were found in the pepper samples and 10 identified mixed infection types were found in the tomato samples. Among the infected samples, Tomato spotted wilt orthotospovirus (TSWV) was the most common virus, with a detection rate of about 20.0% followed by Pepper vein yellows virus (PeVYV, 13.0%). This survey revealed for the first time that pepper is a natural host of Tobacco vein distorting virus (TVDV) worldwide and tomato is a natural host of Potato leafroll virus (PLRV) in China. PeVYV, Tobacco mild green mosaic virus (TMGMV) and Wild tomato mosaic virus (WTMV) were first time found in pepper and Tomato mottle mosaic virus (ToMMV) and Chilli veinal mottle virus (ChiVMV) were first time found in tomato in Yunnan Province. Finally, the virus incidences were higher in Kunming, Yuxi, Chuxiong, and Honghe region than other regions.

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rSodC is a potential antigen to diagnose Corynebacterium pseudotuberculosis by enzyme-linked immunoassay

AMB Express ◽

10.1186/s13568-020-01125-0 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Antonio Pedro Fróes de Farias ◽

José Tadeu Raynal Rocha Filho ◽

Silvana Beutinger Marchioro ◽

Luan Santana Moreira ◽

Andressa Souza Marques ◽

...

Keyword(s):

Sensitivity And Specificity ◽

Recombinant Proteins ◽

Corynebacterium Pseudotuberculosis ◽

Semiarid Region ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Serological Tests ◽

Control Programs ◽

Epidemiological Surveys ◽

Purified Antigen

Abstract Caseous lymphadenitis (CL) is a chronic infectious disease that affects sheep and goats. Many serological tests have been developed to detect the disease; one of the most widely used is the enzyme-linked immunosorbent assay (ELISA), due to its advantages, which include acceptable cost-effectiveness, applicability, sensitivity and specificity. ELISA formulations using recombinant proteins can exhibit significant sensitivity and specificity when using a single purified antigen. DTxR, Trx, TrxR, LexA, SodC, SpaC, NanH, and PknG recombinant proteins can be considered target proteins for ELISA development due to its extracellular or on the cell surface location, which allows a better recognition by the immune system. Therefore, the objectives of this study were to evaluate the antigenic reactivity of Corynebacterium pseudotuberculosis recombinant proteins in goat and sheep serum. Of eight proteins evaluated, rSodC was selected for validation assays with small ruminant serum samples from the semiarid region of the state of Bahia, Brazil. Validation assays with goat serum samples showed that ELISA-rSodC presented sensitivity and specificity of 96% and 94%, respectively. Validation assays with sheep serum showed that ELISA-rSodC exhibited sensitivity and specificity of 95% and 98%, respectively. Analysis of 756 field serum samples showed that rSodC identified 95 positive samples (23%) in goats and 75 positive samples (21%) in sheep. The ELISA with recombinant SodC protein developed in this study discriminated positive and negative serum samples with high levels of sensitivity and specificity. This formulation is promising for epidemiological surveys and CL control programs. Trial registration AEC No 4958051018. 12/18/2018, retrospectively registered

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Natural Infection of Field-Grown Borage (Borago officinalis) by Alfalfa mosaic virus in Spain

Plant Disease ◽

10.1094/pdis.2002.86.6.698a ◽

2002 ◽

Vol 86 (6) ◽

pp. 698-698 ◽

Cited By ~ 2

Author(s):

C. Mallor ◽

M. Luis-Arteaga ◽

M. A. Cambra ◽

S. Fernández-Cavada

Keyword(s):

Mosaic Virus ◽

Indicator Species ◽

Natural Infection ◽

Alfalfa Mosaic Virus ◽

Solanum Melongena ◽

Enzyme Linked Immunosorbent Assay ◽

Human Consumption ◽

Vegetable Crops ◽

Borago Officinalis ◽

Local Lesions

Alfalfa mosaic virus (AMV) has a wide host range and is distributed throughout the world. It causes disease in several vegetable crops, including bean, celery, lettuce, pea, pepper, and tomato (1). In Spain, it has been found naturally infecting alfalfa, pepper, and tomato. During the autumn of 1999, in the area of Zaragoza (northeastern Spain), several plants expressing foliar yellow mosaic symptoms were observed in borage grown for human consumption in open field plots. The commercial value of the symptomatic plants was greatly reduced. The symptoms were similar to those previously obtained in greenhouse-grown borage plants mechanically inoculated with three tomato isolates of AMV (2). The following indicator species, including virus-free borage plants, were mechanically inoculated with sap from leaves of symptomatic borage plants, and reactions were recorded: chlorotic and necrotic local lesions on Tetragonia expansa and Vigna unguiculata; chlorotic local lesions and systemic mosaic on Chenopodium quinoa, C. amaranticolor, Cucumis sativus ‘Marketmore’, Gomphrena globosa, and Nicotiana glutinosa; systemic mosaic, sometimes associated with localized reactions, on Ocimum basilicum, Capsicum annuum ‘Doux des Landes’ and ‘Yolo Wonder’, N. benthamiana, N. clevelandii, N. rustica, N. sylvestris, N. tabacum ‘Paraguay’ and ‘Xanthi nc’, Petunia hybrida, Physalis floridana, and Solanum melongena ‘Cerna krazavitska’. The reactions are in agreement with the indicator host reactions described for AMV (1). Symptoms on virus-free borage plants mechanically inoculated with sap from symptomatic borage and from inoculated C. amaranticolor and O. basilicum experimental hosts were similar to those observed in naturally infected borage. Positive serological reactions (A405 values more than three times greater than those of the negative controls) in double-antibody sandwich enzyme-linked immunosorbent assay using commercially prepared antiserum against AMV (Agdia, Inc., Elkhart, IN) were obtained with extracts of naturally infected borage leaves and with systemically infected indicator species. Alfalfa plots located in the vicinity of the symptomatic borage plants could be the source of virus for borage infections. To our knowledge, this is the first report of natural AMV infection in Borago spp. References: (1) E. M. J. Jaspars and L. Bos. CMI/AAB. No. 229, 1980. (2) M. Luis-Arteaga and J. M. Alvarez. Inf. Téc. Econ. Agr. 92:70, 1996.

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Occurrence of a New Pathotype of Lettuce mosaic virus on Lettuce in Brazil

Plant Disease ◽

10.1094/pdis.2000.84.4.490a ◽

2000 ◽

Vol 84 (4) ◽

pp. 490-490 ◽

Cited By ~ 7

Author(s):

O. Stangarlin ◽

M. A. Pavan ◽

N. da Silva

Keyword(s):

Electron Microscopy ◽

Mosaic Virus ◽

Mixed Infections ◽

Lettuce Mosaic Virus ◽

Chenopodium Amaranticolor ◽

Serological Tests ◽

Virus Identification ◽

Plant Pathol ◽

Differential Hosts ◽

Important Disease

Since 1970 lettuce mosaic has not been an important disease of lettuce in Brazil, due to the growing of cultivars that contain gene g, derived from cv. Gallega de Invierno, for tolerance. Recently, however, a widespread, serious outbreak of lettuce mosaic occurred in the State of Sao Paulo. Both lettuce cultivars that lack and those that contain gene g have been affected, suggesting the emergence of a new pathotype of Lettuce mosaic virus (LMV). Commercial lettuce fields were visited, and 20 samples were collected for virus identification by bioassay on differential hosts, serological tests, and electron microscopy. Of these, 12 were infected only by LMV, and 8 contained mixed infections of LMV and a possible new sequivirus, Lettuce mottle virus. Seedlings of susceptible cv. White Boston were inoculated with LMV from tolerant cultivars, and plants were allowed to flower. Seeds were collected and sown, and seedlings with mosaic symptoms were recovered. Twenty monolesional isolates, obtained by passing the virus three times through Chenopodium amaranticolor, were inoculated individually on lettuce differential cvs. Salinas and White Boston (susceptible) and cvs. Salinas 88, Vanguard 75, Ithaca, Malika, and Gallega de Invierno (tolerant) to previously described pathotypes of LMV (1). Considering the susceptibility of all test differentials, we concluded that a LMV pathotype IV exists that has overcome tolerance conferred by genes m and g and is responsible for the new outbreak of LMV in Brazil. This is the first report of LMV-IV in Latin America. Reference: (1) D. A. C. Pink et al. Plant Pathol. 41:5, 1992.

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rSodC is a potential antigen to diagnose Corynebacterium pseudotuberculosis by enzyme-linked immunoassay

10.21203/rs.3.rs-54005/v2 ◽

2020 ◽

Author(s):

Antonio Pedro Fróes de Farias ◽

José Tadeu Raynal Rocha Filho ◽

Silvana Beutinger Marchioro ◽

Luan Santana Moreira ◽

Andressa Souza Marques ◽

...

Keyword(s):

Sensitivity And Specificity ◽

Recombinant Proteins ◽

Corynebacterium Pseudotuberculosis ◽

Semiarid Region ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Serological Tests ◽

Control Programs ◽

Epidemiological Surveys ◽

Purified Antigen

Abstract Caseous lymphadenitis (CL) is a chronic infectious disease that affects sheep and goats. Many serological tests have been developed to detect the disease; one of the most widely used is the enzyme-linked immunosorbent assay (ELISA), due to its advantages. ELISA formulations using recombinant proteins can exhibit significant sensitivity and specificity when using a single purified antigen. DTxR, Trx, TrxR, LexA, SodC, SpaC, NanH, and PknG recombinant proteins can be considered target proteins for ELISA development. Therefore, the objectives of this study were to evaluate the antigenic reactivity of Corynebacterium pseudotuberculosis recombinant proteins in goat and sheep serum. Of eight proteins evaluated, rSodC was selected for validation assays with small ruminant serum samples from the semiarid region of the state of Bahia, Brazil. Validation assays with goat serum samples showed that rSodC presented sensitivity and specificity of 96% and 94%, respectively. Validation assays with sheep serum showed that recombinant SodC exhibited sensitivity and specificity of 95% and 98%, respectively. Analysis of 756 field serum samples showed that rSodC identified 95 positive samples (23%) in goats and 75 positive samples (21%) in sheep. The ELISA with recombinant SodC protein developed in this study discriminated positive and negative serum samples with high levels of sensitivity and specificity. This formulation is promising for epidemiological surveys and CL control programs.

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