Sugar composition and serological specificity of Erwinia carotovora lipopolysaccharides

1985 ◽  
Vol 31 (7) ◽  
pp. 583-586 ◽  
Author(s):  
S. H. De Boer ◽  
J. J. Bradshaw-Rouse ◽  
L. Sequeira ◽  
M. E. McNaughton
Keyword(s):  
Immunosorbent Assay ◽  
Type Strain ◽  
Erwinia Carotovora ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sugar Composition ◽  
Whole Cell ◽  
Precipitin Line ◽  
Whole Cells ◽  
Cross Reactions ◽  
Serological Specificity

Sugar composition and serological specificity of the lipopolysaccharides (LPS) purified from 16 Erwinia carotovora strains in six different serogroups were determined. All the LPS preparations contained 2-keto-3-deoxyoctonoic acid, glucosamine, heptose, and glucose, and most contained galactose. Either rhamnose or fucose was present in LPS from 12 of the strains, and the presence or absence of these deoxy sugars was consistent for all strains within a serogroup. LPS from two strains contained mannose. One unidentified sugar was present in all LPS preparations, but six other unidentified sugars varied in different LPS preparations. All LPS preparations reacted in an enzyme-linked, immunosorbent assay (ELISA) with antisera produced against whole cells of a type strain in the same serogroup as the strain from which the LPS was extracted. Several cross-reactions among strains that previously were observed in immunodiffusion tests with whole-cell preparations were also observed in ELISA with purified LPS. Some of the LPS preparations also reacted in immunodiffusion with a precipitin line of identity with whole-cell preparations. The results indicate that E. carotovora serogroups probably are based on the LPS 0-antigen.

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay for Serotyping Ureaplasma urealyticum Strains Using Monoclonal Antibodies

2001 ◽  
Vol 8 (1) ◽  
pp. 52-57 ◽  
Author(s):  
Fedoua Echahidi ◽  
Gaëtan Muyldermans ◽  
Sabine Lauwers ◽  
Anne Naessens
Keyword(s):  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
Ureaplasma Urealyticum ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
Cross Reactions ◽  
Antigenic Preparation ◽  
Negative Controls ◽  
Reference Strains

ABSTRACT Ureaplasma urealyticum comprises 14 serotypes. The existing serotyping methods all use polyclonal antibodies. These methods are time-consuming and labor-intensive, and they cannot always be performed on primary isolates; in addition, the results are difficult to interpret. We developed a new enzyme-linked immunosorbent assay (ELISA) method using serotype-specific monoclonal antibodies (MAbs) to enable the serotyping of U. urealyticum isolates from primary broth cultures. Each of the 14 serotype reference strains was tested against 14 selected MAbs. Homologous reactions were very strong, while heterologous reactions were negligible. Three cross-reactions were observed: MAb 5 cross-reacted with serotype 2, MAb 14 cross-reacted with serotype 3, and MAb 8 cross-reacted with serotype 13. Despite the cross-reactions observed, all the serotype reference strains ofU. urealyticum could be identified and differentiated using a combination of MAbs. Reproducibility was analyzed with a fractionated antigenic preparation and with several freshly prepared antigens of the same strain. No significant interrun variation was found with the fractionated antigen, but significant variations in optical density (OD) values were found when freshly prepared antigens were tested. However, the variation in OD values did not influence the overall interpretation of the ELISA: reactions with homologous MAbs were always prominent compared to those of the negative controls. This newly developed ELISA using MAbs seems promising for serotyping of U. urealyticum clinical isolates.

scholarly journals Vaccination in Nile tilapia broodstock with whole cell vaccine and disease resistance in its fry against Aeromonas hydrophila

2017 ◽  
Vol 16 (2) ◽  
pp. 268 ◽  
Author(s):  
Sukenda Sukenda ◽  
Odang Carman ◽  
Rahman Rahman ◽  
Dendi Hidayatullah ◽  
Nurfitriani Siti Yumaidawati
Keyword(s):  
Disease Resistance ◽  
Immunosorbent Assay ◽  
Antibody Level ◽  
Aeromonas Hydrophila ◽  
Nile Tilapia ◽  
Cell Vaccine ◽  
Enzyme Linked Immunosorbent Assay ◽  
Whole Cell ◽  
Whole Cell Vaccine ◽  
Phosphate Buffered Saline

<p class="NoParagraphStyle"><strong>ABSTRACT</strong></p><p class="NoParagraphStyle"><strong> </strong></p><p class="NoParagraphStyle">The aim of this study was to analyze the effectivity of vaccination in Nile tilapia broodstock with whole cell vaccine and disease resistance in fry tilapia against <em>Aeromonas hydrophila</em>. Tilapia Nirwana strain that used for this had average body weight of 185±13.23 g and were maintained in ponds sizing of (2.5×2.5×1 m<sup>3</sup>). Vaccinations that has been done through intraperitoneal injection using dose of 0.1 mL/fish, meanwhile the fish for control was injected by phosphate buffered saline (PBS). This study used complete randomized design with two treatments and three replications. Antibody level was measured by using indirect enzyme-linked immunosorbent assay (ELISA) method in the broodstock, egg, and fry.  Challenge test in fry tilapia performed at the age of 5, 10, and 15 days. The results showed that vaccination in tilapia broodstock delivered a significant antibody level in broodstock, eggs, and fry (P&lt;0.05) compared to the control. Relative percent survival of offspring at 5, 10, and 15 days were 78.26%, 70.59%, and 65.52%, respectively.  As a conclusion, vaccination in tilapia broodstock was effective to improve specific and non-specific immunity, and protect fry tilapia from <em>A. hydrophila</em> infection through maternal immunity.</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Keywords: vaccination, antibody, maternal immunity, tilapia, <em>Aeromonas hydrophila</em></p><p class="NoParagraphStyle"><strong> </strong></p><p class="NoParagraphStyle"><strong> </strong></p><p class="NoParagraphStyle"><strong>ABSTRAK</strong><strong></strong></p><p class="NoParagraphStyle"><strong> </strong></p><p class="NoParagraphStyle">Penelitian ini bertujuan untuk menganalisis efikasi vaksinasi pada induk nila dengan vaksin sel utuh dan ketahanan benih yang dihasilkan terhadap <em>Aeromonas hydrophila</em>. Ikan nila stain Nirwana yang digunakan dalam penelitian memiliki bobot rata-rata 185±13,23 g dan ikan dipelihara dalam kolam (2,5×2,5×1 m<sup>3</sup>). vaksinasi dilakukan melalui penyuntikan intraperitoneal dengan dosis 0,1 mL/ikan, sementara itu ikan kontrol disuntik dengan <em>phosphate buffered saline</em> (PBS). Penelitian ini menggunakan rancangan acak lengkap dengan dua perlakuan dan tiga ulangan. Tingkat antibodi diukur dengan menggunakan metode<em> indirect enzyme-linked immunosorbent assay</em> (ELISA) pada induk, telur dan benih. Uji tantang pada benih dilakukan pada umur 5, 10, dan 15 hari. Hasil penelitian menunjukan bahwa vaksinasi pada induk nila secara signifikan dapat meningkatkan level antibodi pada induk, telur, dan benih (P&lt;0,05) dibandingkan dengan kontrol. Kelangsungan hidup relatif pada benih berumur 5, 10, dan 15 hari masing-masing adalah 78,26%; 70,59%; dan 65,52%. Sebagai kesimpulan vaksinasi pada induk nila efektif dalam memperbaiki imunitas spesifik dan non spesifik serta melindungi benih dari infeksi <em>A. hydrophila</em> melalui imunitas maternal.</p><p class="NoParagraphStyle"> </p><p>Kata kunci: vaksinasi, antibodi, imunitas maternal, ikan nila, <em>Aeromonas hydrophila</em></p>

scholarly journals Enzyme-Linked Immunosorbent Assay for Salmonella Serology using Lipopolysaccharide Antigen

1995 ◽  
Vol 7 (4) ◽  
pp. 481-487 ◽  
Author(s):  
Bradford P. Smith ◽  
George W. Dilling ◽  
John K. House ◽  
Hans Konrad ◽  
Nadia Moore
Keyword(s):  
Immunosorbent Assay ◽  
Epidemiologic Studies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cross Reactions ◽  
Vaccine Responses ◽  
O Antigen ◽  
Specific Igg ◽  
O Antigens ◽  
Somatic Antigens ◽  
Factor Antigen

Enzyme-linked immunosorbent assay (ELISA) using Salmonella lipopolysaccharide (LPS) to measure specific IgG titers in cattle has proven useful. Serology can be used to assess vaccine responses and infection rates, to detect carriers, and to aid in epidemiologic studies. The objective of this study was to assess cross-reactions using sera from cattle vaccinated with different Salmonella serogroups. ELISA plates using lipopolysaccharide from serogroup B, C1, C3, D1 or E1 as the plate antigens were tested. LPS was extracted from Salmonella typhimurium (Serogroup B; somatic antigens 01, 4, 12), S. montevideo (C1; 06, 7), S. kentucky (C3; 08, 20), S. dublin (D1; 01, 9, 12) and S. anatum (E1; 03, 10) using the Westphal method. Fifteen cows were found to be seronegative for all 5 of these serogroup antigens. Each cow was then vaccinated 3 times at 2-week intervals with a killed Salmonella bacterin. The 15 different serotypes used for vaccination were chosen to represent a wide array of Salmonella serogroups with a wide array of somatic “O” antigens expressed, including somatic antigens 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 18, 19, 20, 22, 23, 25, and 27. With each antigen tested, the highest ELISA titers were seen with sera from cattle vaccinated with homologous O antigens, indicating that reactions were highly O antigen-specific. Some cross reactions between serogroups sharing one O factor antigen were found; these titers were lower than those found with homologous serogroups sharing 2 or more antigens. Only serum from the cow vaccinated with S. anatum (group E; antigens 03, 10) cross-reacted at a low titer with group C1 (O somatic antigens 6, 7) and D1 (O somatic antigens 1, 9, 12) plate antigens, with which no somatic antigens were shared. We conclude from these results that Salmonella serology using LPS antigens is highly O antigen-specific and predictable.

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