Immunoblot reactivity of polyclonal and monoclonal antibodies with periplasmic flagellar proteins FlaA1 and FlaB of porcine Serpulina species

Fisher, L N; Duhamel, G; Westerman, R B; Mathiesen, Michelle R.

doi:10.1128/cdli.4.4.400-404.1997

Cited by 12 publications

(4 citation statements)

References 30 publications

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“…However, flagellar proteins are highly conserved among spirochete species and genera, therefore false-positive results due to cross-reactivity are always a potential problem if these proteins are used for serologic tests (Jensen, 1997). This is consistent with polyclonal and monoclonal antibody reagents raised against B. hyodysenteriae cross-reacting with B. pilosicoli FlaA1 and FlaB by immunoblot (Fisher et al, 1997).…”

Section: Flagellar Proteinssupporting

confidence: 87%

Serologic detection ofBrachyspira (Serpulina) hyodysenteriaeinfections

Hampson

2001

Anim. Health. Res. Rev.

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Swine dysentery (SD) caused by the intestinal spirochete Brachyspira hyodysenteriae is an economically important disease in pig-producing countries throughout the world. To date, no specific serologic assay is commercially available for the diagnosis of pigs with SD. Several serologic techniques have been identified in the past; however, these tests have all used either whole-cell proteins or lipopolysaccharide (LPS) as the antigen. Whole-cell antigens are plagued with false-positive reactions due to cross-reactivity with common proteins shared with other spirochetes. LPS antigens produce fewer false-positives; however, false-negatives may result due to LPS components being serogroup-specific. Generally, these techniques are useful for detecting infected herds, but are unreliable for the detection of individual infected pigs. In order to develop improved serologic tests it will be necessary to identify suitable diagnostic antigens, in particular immunogenic cell-surface structures which are specific to B. hyodysenteriae but common amongst different strains of the species. Recently, we identified and cloned a 30-kDa outer membrane lipoprotein (BmpB) which is specific to B. hyodysenteriae and is recognized by experimentally and naturally infected pigs. In this review we summarize the available serologic tests for SD, and speculate on the use of recombinant BmpB as an antigen for future development of an improved serologic test for SD diagnosis.

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Section: Flagellar Proteinssupporting

confidence: 87%

Serologic detection ofBrachyspira (Serpulina) hyodysenteriaeinfections

Hampson

2001

Anim. Health. Res. Rev.

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“…Koopman and coworkers (24) and Li and coworkers (30) reported that PF protein FlaA1 has the greatest variability among the five PF proteins of intestinal spirochetes. We have shown that the PF FlaA1 protein is antigenically similar among porcine Serpulina species, as demonstrated by the immunoblot reactivity of an FlaA1-specific rabbit antiserum to the 44-kDa PF protein of intestinal spirochetes (13,30). However, Koopman and coworkers (24) suggested that there was polymorphism of the PF flaA1 gene among porcine S. hyodysenteriae isolates B204 and C5, S. innocens isolate B256, human B. aalborgi, and 24 nonclassified WBHIS isolated from humans (n ϭ 10), dogs (n ϭ 5), rats (n ϭ 4), mice (n ϭ 4), and a chicken.…”

Section: Discussionmentioning

confidence: 87%

Restriction fragment length polymorphism of the periplasmic flagellar flaA1 gene of Serpulina species

Fisher

Mathiesen

Duhamel

1997

Clin Diagn Lab Immunol

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Forty-one reference and field isolates of intestinal spirochetes representing Serpulina hyodysenteriae, Serpulina innocens, Serpulina pilosicoli, Brachyspira aalborgi, and nonclassified weakly beta-hemolytic intestinal spirochetes were compared by restriction fragment length polymorphism (RFLP) of the periplasmic flagellar (PF) flaA1 gene. Six genetically distinct groups (I through VI), each with a unique RFLP fingerprint pattern, were identified by Southern blotting analysis of EcoRV chromosomal DNA digests with a PCR-amplified digoxigenin-labeled 1-kb fragment of the S. hyodysenteriae isolate B78 PF flaA1 gene. The RFLP fingerprint patterns corresponded to known DNA homology differences between Serpulina species and to provisionally designated species described previously by using phenotypic and genotypic classification schemes. RFLP fingerprinting of the PF flaA1 gene provides a relatively simple genotypic method for identification of intestinal spirochetes without the use of radioisotopes.

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“…Flagellar proteins are major immunoreactive proteins in B. hyodysenteriae (Kent et al, 1989 ). It has been reported that FlaA from S. hyodysenteriae, S. innocens , and S. pilosicoli was recognized by rabbit polyclonal and murine monoclonal antibodies produced against S. hyodysenteriae lysates (Fisher et al, 1997 ). However, no data were previously available on the antigenicity of B. pilosicoli FlaA.…”

Section: Discussionmentioning

confidence: 99%

Brachyspira hyodysenteriae and B. pilosicoli Proteins Recognized by Sera of Challenged Pigs

et al. 2017

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The spirochetes Brachyspira hyodysenteriae and B. pilosicoli are pig intestinal pathogens that are the causative agents of swine dysentery (SD) and porcine intestinal spirochaetosis (PIS), respectively. Although some inactivated bacterin and recombinant vaccines have been explored as prophylactic treatments against these species, no effective vaccine is yet available. Immunoproteomics approaches hold the potential for the identification of new, suitable candidates for subunit vaccines against SD and PIS. These strategies take into account the gene products actually expressed and present in the cells, and thus susceptible of being targets of immune recognition. In this context, we have analyzed the immunogenic pattern of two B. pilosicoli porcine isolates (the Spanish farm isolate OLA9 and the commercial P43/6/78 strain) and one B. hyodysenteriae isolate (the Spanish farm V1). The proteins from the Brachyspira lysates were fractionated by preparative isoelectric focusing, and the fractions were analyzed by Western blot with hyperimmune sera from challenged pigs. Of the 28 challenge-specific immunoreactive bands detected, 21 were identified as single proteins by MS, while the other 7 were shown to contain several major proteins. None of these proteins were detected in the control immunoreactive bands. The proteins identified included 11 from B. hyodysenteriae and 28 from the two B. pilosicoli strains. Eight proteins were common to the B. pilosicoli strains (i.e., elongation factor G, aspartyl-tRNA synthase, biotin lipoyl, TmpB outer membrane protein, flagellar protein FlaA, enolase, PEPCK, and VspD), and enolase and PEPCK were common to both species. Many of the identified proteins were flagellar proteins or predicted to be located on the cell surface and some of them had been previously described as antigenic or as bacterial virulence factors. Here we report on the identification and semiquantitative data of these immunoreactive proteins which constitute a unique antigen collection from these bacteria.

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Immunoblot reactivity of polyclonal and monoclonal antibodies with periplasmic flagellar proteins FlaA1 and FlaB of porcine Serpulina species

Cited by 12 publications

References 30 publications

Serologic detection ofBrachyspira (Serpulina) hyodysenteriaeinfections

Serologic detection ofBrachyspira (Serpulina) hyodysenteriaeinfections

Restriction fragment length polymorphism of the periplasmic flagellar flaA1 gene of Serpulina species

Brachyspira hyodysenteriae and B. pilosicoli Proteins Recognized by Sera of Challenged Pigs

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