Showing papers on "Hemagglutination published in 2005"
TL;DR: The assay for natural antibody-mediated complement activation and red blood cell agglutination described here, requiring approximately 100 microl of blood, is highly repeatable and can be used to compare constitutive innate humoral immunity among species and with respect to age, sex, and experimental treatments within populations.
Abstract: Methods to assess immunocompetence requiring only a single sample are useful in comparative studies where practical considerations prevent holding or recapturing individuals. The assay for natural antibody-mediated complement activation and red blood cell agglutination described here, requiring ∼100 μl of blood, is highly repeatable. The effects of complement deactivation, 2-mercaptoethanol (2-ME), age, and lipopolysaccharide (LPS)-induced sickness response were examined to validate comparisons among diverse avian species. Complement deactivation by heating significantly reduces lysis and treatment with 2-ME reduces both lysis and agglutination. Lysis and agglutination both increase with age in chickens; LPS treatment does not influence these variables in 11-week-old chickens. In a comparison of 11 species, both lysis (0.0–5.3 titers) and agglutination (1.8–8.0 titers) vary significantly among species. Accordingly, this assay can be used to compare constitutive innate humoral immunity among species and with respect to age, sex, and experimental treatments within populations.
388 citations
TL;DR: In most of the parameters, L. rohita showed the highest value, possibly indicating its more natural resistance compared with the other two species, as well as indicating the immunomodulatory substances for fish farming as markers of pollution and disease resistance.
Abstract: Summary The non-specific immune parameters are useful to determine the health status of fish and to evaluate the immunomodulatory substances for fish farming as markers of pollution and disease resistance. Some of the important parameters, viz. superoxide production by neutrophils through nitroblue tetrazolium (NBT) assay; haemagglutination (HA), haemolysin (HLY) and bacterial agglutination titres; myeloperoxidase (MPO), and lysozyme activities, and alternative complement levels in serum of the juveniles of three Indian major carp species (Cirrhinus mrigala, Catla catla and Labeo rohita) were measured to establish their physiological normal range. A wide variation among the individuals within a species in the ranges of the most of the immune parameters was recorded. Significantly higher levels in the mean values of HA, HLY, bacterial agglutination titres; superoxide production by neutrophils in nitroblue tetrazolium assay; serum MPO and lysozyme activities, i.e. 371.20, 4.60, 18.80, 0.40, 0.62 and 6.55 l gm l )1 , respectively, were obtained in L. rohita except a much lower alternative haemolytic complement activity (29.06 units ml )1 ) compared with the other two species. In most of the parameters, L. rohita showed the highest value, possibly indicating its more natural resistance compared with the other two species.
180 citations
TL;DR: Five laboratory tests for diagnosis of canine parvovirus type 2 (CPV-2) infection were employed to test 89 faecal samples collected from dogs with diarrhoea, finding the best correlation was found between conventional and real-time PCR.
176 citations
TL;DR: ABO‐FACS is a valid method to quantify anti‐A/B IgM, IgG and IgG subclasses to human A or B red blood cells and opens the possibility of isotype‐specific monitoring of anti‐ A/B antibodies levels after ABO‐incompatible solid organ and stem cell transplantation.
Abstract: Several methods to detect anti-A/B antibodies based on haemagglutination and haemolysis have been described. These methods measure predominantly anti-A/B immunoglobulin (Ig)M, whereas anti-A/B IgG and IgG subclasses are less well examined. We established a flow cytometry method (ABO-fluorescence-activated cell sorting; ABO-FACS) to quantify binding of anti-A/B IgM, IgG and IgG subclasses to human A or B red blood cells. Anti-A/B IgM were present in the majority of 120 blood donors, as expected from blood group typing. The sensitivity and specificity of anti-A/B IgM to predict the blood group was 93% and 96% respectively. Anti-A/B IgG was found in 34/38 blood group O samples (89%). Anti-B IgG in blood group A or anti-A IgG in blood group B was present in 4/28 (14%) and 1/28 (4%) samples, respectively, and absent in 26 AB sera. IgG2 was the predominant IgG subclass. The correlation of anti-A/B IgM and IgG in the ABO-FACS with haemagglutination titres was 0.870 and 0.783, respectively (n = 240; P < 0.001) whereas the comparison of ABO-FACS with ABO-enzyme-linked immunosorbent assay was less significant. In conclusion, ABO-FACS is a valid method to quantify anti-A/B IgM, IgG and IgG subclasses. It opens the possibility of isotype-specific monitoring of anti-A/B antibodies levels after ABO-incompatible solid organ and stem cell transplantation.
108 citations
TL;DR: It is suggested that GT1b gangliosides react specifically with the virus protein responsible for membrane fusion and thus are involved in virus penetration and delivery of the virus genome to the nuclei.
Abstract: The ganglioside composition of Ehrlich ascites carcinoma (EAC) cells and the role of the individual gangliosides in binding and penetration into the cell of influenza virus were determined. EAC gangliosides identical with or close to GM3, GM2, GM1, GT1a and GT1b were characterized by thin-layer chromarography, compositional analyses, methylation analysis and mass-spectrometry. The ganglioside uptake capacity of native and neuraminidase-treated EAC cells was studied with tritium-labeled gangliosides of definite structure and the binding of influenza virus to cells was determinated by using [3H]uridine-labeled virus and by hemagglutination studies. Treatment of the cells with Vibrio cholerae neuraminidase largely decreased binding of the virus. Exogenous gangliosides with a terminal galactose unit or a penultimate galactose masked by neuraminic acid were able to restore the virus-binding capacity of neuraminidase-treated cells, however, the main ganglioside of EAC cells, GM2, which carbohydrate chain is terminated by N-acetylgalactosamine, was completely ineffective. The common carbohydrate sequence of the ganghosides showing binding activity
is proposed to be the main recognition structure of the influenza virus receptor on the surface of EAC cells.
Penetration of labeled influenza virus into the nuclei of EAC cells was evaluated by measuring the radioactivity of the nuclei of neuraminidase-treated ganglioside-loaded cells after exposition to the labeled virus. Of all gangliosides tested only trisialogangliosides of the GT1b type were able to induce increased entry of the virus into the cells and accumulation of its radioactive component into the nuclei. It is suggested that GT1b gangliosides react specifically with the virus protein responsible for membrane fusion (apparently the hemagglutinin HA2 subunit) and thus are involved in virus penetration and delivery of the virus genome to the nuclei.
105 citations
TL;DR: The A/Turkey/OH/313053/04 (H3N2) isolate isolated from turkeys in this study was classified as a low pathogenic avian influenza A virus because it only caused a drop in egg production with minor other clinical signs and no mortality.
Abstract: Five 34-wk-old turkey breeder layer flocks in separate houses of 2550 birds each in a single farm in Ohio experienced a drop in egg production from late January to early February 2004. Tracheal swabs (n = 60), cloacal swabs (n = 50), and convalescent sera (n = 110) from the flocks were submitted to the laboratory for diagnostics. Virus isolation was attempted in specific-pathogen free embryonating chicken eggs and Vero and MDCK cells. Virus characterization was performed using agar gel immunodiffusion, the hemagglutination test, the hemagglutination inhibition test, the virus neutralization test, reverse transcription–polymerase chain reaction, sequencing, and phylogenetic analysis. A presumptive influenza virus was successfully propagated and isolated on the first passage in MDCK cells, but initially not in Vero cells or specific-pathogen free chicken embryos. After two passages in MDCK cells, it was possible to propagate the isolate in specific-pathogen free chicken embryos. Preliminary sequenc...
89 citations
TL;DR: Cranberry juice contains high molecular weight materials (NDM) that inhibit bacterial adhesion to host cells as well as the co-aggregation of many oral bacteria, which indicates that NDM may have a therapeutic potential for influenza virus adhesion and infectivity.
82 citations
TL;DR: The interaction between maternally-derived antibodies and canine parvovirus (CPV) infection was evaluated in five groups of pups with a wide range of haemagglutination inhibiting (HI) titres of MDA, and the need for revision of current vaccination programmes for pups is suggested.
70 citations
TL;DR: A lectin isolated and purified from the culture filtrate of the plant pathogenic fungus Macrophomina phaseolina exhibited binding affinity towards neuraminyl oligosaccharides of N-linked glycoproteins, alpha-Neu5Ac-(2-->3)-beta-Gal-(1-->4)-GlcNAc being maximum.
48 citations
TL;DR: It is concluded that specific forms of sialic acid are used as receptor by recent human H3N2 influenza viruses, perhaps involving branched alpha2,6 sIALic acid oralpha2,8 sialIC acid structures on O-linked carbohydrates.
44 citations
TL;DR: It is demonstrated that immunisation with ganglioside-mimicking C. jejuni LOS triggers the production of cross-reactive anti-gangliosid antibodies that recognise epitopes at the nodes of Ranvier in sectioned human nerves.
TL;DR: In this article, the authors used a polymerase chain reaction (PCR) to determine the importance of expression of adhesins in the pathogenic Escherichia coli (APEC) strain and evaluated the presence of genes either previously described or not yet recognized for APEC strain.
TL;DR: Preliminary results showed that the antigen administered within the cell debris fraction of the transformed yeast protected rabbits immunized by the oral route against an intramuscular challenge with 100 LD50 (16,000 hemagglutination units) of homologous virus.
TL;DR: An indirect hemagglutination assay routinely used to detect antibodies to Burkholderia pseudomallei was modified to detect cross-reactivity of antibodies to B. pseudomaliani, B. mallei, and B. thailandensis antigens and it is demonstrated a lack of cross- reactivity between B. Pseudo-Pseudo-Mallei and B Thailandensis.
Abstract: The indirect hemagglutination assay routinely used to detect antibodies to Burkholderia pseudomallei was modified to detect cross-reactivity of antibodies to B. pseudomallei, B. mallei, and B. thailandensis antigens. We demonstrate a lack of cross-reactivity between B. pseudomallei and B. thailandensis but marked cross-reactivity between B. pseudomallei and B. mallei.
TL;DR: CPV-2b was the prevalent type circulating in the State of Rio de Janeiro from 1995 to 2001, and was confirmed by PCR in 5 of these 15 puppies who had received old-type vaccines.
Abstract: In this study, the genomic types of canine parvovirus (CPV) circulating in the State of Rio de Janeiro, Brazil, from 1995 to 2001, were investigated using the polymerase chain reaction assay (PCR). A total of 78 faecal samples from gastroenteritic puppies, confirmed as positive for canine parvovirus by haemagglutination/haemagglutination inhibition tests or virus isolation in cell culture (MDCK), were examined. The viral DNA was extracted from faecal samples using a combination of phenol– chloroform and silica–guanidine thiocyanate methods. PCR was carried out with differential pairs of primers to distinguish the old (CPV-2) and new types of virus (CPv-2a or CPV-2b). Specific amplicons were observed for all samples using the primer pair P2ab, which detects CPV-2a and CPV-2b. Seventy-six from a total of 78 samples (97%) were considered as CPV-2b because of their reaction with the primer pair P2b. Thirty samples (30/78) were from previously vaccinated puppies and in 15 of them the enteritis symptoms began from 1 to 12 days after vaccination. PCR confirmed the infection by wild virus (CPV-2b) in 5 of these 15 puppies who had received old-type vaccines. Our results show that CPV-2b was the prevalent type circulating in the State of Rio de Janeiro from 1995 to 2001.
TL;DR: Seven Peste des petits ruminants viruses and a RPV (vaccine strain) and their respective antiserum raised in rabbits were used for reciprocal cross neutralization test and haemagglutination andHI test employing 0.5% chicken RBC.
TL;DR: The results of the modelling approach indicated that the structure of a variant C-Yoichi HA-33 molecule reveals clear deformation of the beta-trefoil domain responsible for the carbohydrate recognition site, leading to the conclusion that the binding of four HA- 33 molecules is required for haemagglutination activity of botulinum L-TC.
Abstract: Normally, large-sized botulinum toxin complexes (L-TC) of serotype C and D are composed of a single neurotoxin, a single non-toxic non-haemagglutinin, two HA-70 molecules, four HA-33 molecules and four HA-17 molecules that assemble to form a 650 kDa L-TC. The 540 and 610 kDa TC species (designated here as L-TC2 and L-TC3, respectively) were purified in addition to the 650 kDa L-TC from the culture supernatants of serotype D strains (D-4947 and D-CB16) and serotype C strains (C-6814 and C-Yoichi). The 650 kDa L-TC from D-4947, D-CB16 and C-6814 showed haemagglutination and erythrocyte-binding activity, but their L-TC2 and L-TC3 species had only binding activity. In contrast, every TC species from C-Yoichi having the C-terminally truncated variant of HA-33 exhibited neither haemagglutination activity nor erythrocyte-binding activity. Four strain-specific HA-33/HA-17 complexes were isolated from the 650 kDa L-TC of each strain. The 650 kDa HA-hybrid L-TCs were reconstituted by various combinations of isolated HA-33/HA-17 complexes and haemagglutination-negative L-TC2 or L-TC3 from each strain. HA-hybrid 650 kDa L-TC, including at least one HA-33/HA-17 complex derived from C-Yoichi, lost haemagglutination activity, leading to the conclusion that the binding of four HA-33 molecules is required for haemagglutination activity of botulinum L-TC. The results of the modelling approach indicated that the structure of a variant C-Yoichi HA-33 molecule reveals clear deformation of the β-trefoil domain responsible for the carbohydrate recognition site.
TL;DR: The constructed Xeno-mAbs, IgG2 subclass, significantly inhibited hemagglutination of P. gingivalis and its vesicles, and may prove to be useful for the development of passive immunization against periodontal diseases caused by P.
TL;DR: For almost 20 years, the neutralizing-epitope site specific for influenza B virus Victoria group isolates was conserved at the “tip” of the hemagglutinin molecule; however, it was not detected in half of the isolates from the 2002-2003 epidemic in Japan.
Abstract: For almost 20 years, the neutralizing-epitope site specific for influenza B virus Victoria group isolates was conserved at the “tip” of the hemagglutinin molecule; however, it was not detected in half of the isolates from the 2002-2003 epidemic in Japan. Amino acid substitutions (D164E or N165K) were observed at the “tip,” and the epitope was altered. The viral antigenicities were affected, and human antibodies did not substantially inhibit the hemagglutination in the hemagglutination inhibition tests. It is suspected that such variants will be important in future epidemics.
TL;DR: In this study, recombinant anti-F5 scFv antibody fragment produced in E. coli HB2151 reacted with F5 fimbriae in ELISA and Western immunoblot, and prevented haemagglutination induced by the binding of the F5F5 to its natural host receptors on horse red blood cells.
Abstract: Bovine colibacillosis caused by enterotoxigenic Escherichia coli (ETEC) is a worldwide problem. Adhesion of ETEC to intestinal cell receptors mediated by the surface protein F5 fimbriae is the initial step in the establishment of colibacillosis. Prevention of ETEC F5(+) adhesion to enterocytes protects newborn calves against collibacillosis. On the enterocytes, the F5 fimbriae bind to a ganglioside that is also found on horse red blood cells. Thus, the presence of F5 fimbriae induces haemagglutination, which is useful as an indicator in a functional assay system. In this study, recombinant anti-F5 scFv antibody fragment produced in E. coli HB2151 reacted with F5 fimbriae in ELISA and Western immunoblot, and prevented haemagglutination induced by the binding of the F5 fimbriae to its natural host receptors on horse red blood cells. Given the ease with which recombinant antibodies can be mass-produced, the presently described scFv may hold promise as a prophylactic agent for colibacillosis.
01 Jan 2005
TL;DR: The experimental results indicate that compound 4 not only inhibits the attachment of virus to the cell surface receptor but also disturbs the release of the progeny viruses from infected cells by inhibiting both hemagglutinin and sialidase of the influenza viruses.
Abstract: The compound Neu5Ac3αF-DSPE (4), in which the C-3 position was modified with an axial fluorine atom, inhibited the catalytic hydrolysis of influenza virus sialidase and the binding activity of hemagglutinin. The inhibitory activities to sialidases were independent of virus isolates examined.With the positive results obtained for inhibition of hemagglutination and hemolysis induced by A/Aichi/2/68 virus,the inhibitory effect of Neu5Ac3αFDSPE (4) against MDCK cells was examined, and it was found that 4 inhibits the viral infection with IC50 value of 5.6 μM based on the cytopathic effects. The experimental results indicate that compound 4 not only inhibits the attachment of virus to the cell surface receptor but also disturbs the release of the progeny viruses from infected cells by inhibiting both hemagglutinin and sialidase of the influenza viruses.The study suggested that the compound is a new class of bifunctional drug candidates for the future chemotherapy of influenza.
TL;DR: The high percentile (35,9%) of seropositive animals found in this study evidenced its importance as infectious sources of the Influenza virus, subtype H3N8, to the national equine flock.
Abstract: The Influenza virus type A, subtype H3N8, is the etiological agent of the Equine Influenza, responsible for several epidemics and endemic respiratory diseases in world level, besides in the Rio de Janeiro State, Brazil. The objective of this work was to evaluate the role of errant equids, as infectious sources of the Influenza virus, subtype H3N8. The survey was performed from the research of specific antibodies for this virus in 1106 sera analyzed by the inhibition of the hemagglutination test. The high percentile (35,9%) of seropositive animals found in this study evidenced its importance as infectious sources of the Influenza virus, subtype H3N8, to the national equine flock.
TL;DR: It appears that elution time could form a basis for rough characterization of isolates of Newcastle disease virus into the three major strains.
Abstract: Elution time of velogenic, mesogenic and lentogenic strains of Newcastle disease virus was determined. The differences in their elution time were also calculated. Four samples, each of a velogenic strain (VGF2), a mesogenic strain (Komarov) and a lentogenic strain (LaSota) were used for hemagglutination test with 0.6% chicken red blood cells. The time it took for wells of the end hemagglutination points (highest dilution that gave agglutination) to elute was recorded as elution time for each sample. The mean elution time of the three strains of Newcastle disease virus differed significantly (p < 0.05). The velogenic strain gave the highest mean elution time of 118 min, followed by the mesogenic strain with 59 min and the lentogenic strain with 25 min. Based on this result it appears that elution time could form a basis for rough characterization of isolates of Newcastle disease virus into the three major strains.
TL;DR: The data supported that the D antigen is the immunodominant component of the Rh system as indicated by the in vitro and in vivo profiles of Rh specificities in the authors' alloimmunized subjects.
Patent•
21 Sep 2005
TL;DR: In this article, a Pseudomonas aeruginosa mannose sensitive haemagglutination pilus strain is characterized by the mannoses sensitive haemaaggluttination pilu strain.
Abstract: The invention relates to a Pseudomonas aeruginosa mannose sensitive haemagglutination pilus strain, which is characterized by the mannose sensitive haemagglutination pilus strain The invention also relates to the culture, treatment, usage and process of using, as well as the pharmaceutical product prepared therefrom
TL;DR: The presence of cations, especially zinc, copper and cerium, increased attachment of human serum proteins to both bacterial species and inhibited adhesion of both species to buccal epithelial cells and hemagglutination.
Abstract: This study investigated the mechanism of protein attachment to the surface of the putative periodontal pathogens Prevotella intermedia and Prevotella nigrescens in artificial gingival crevicular fluid, and ways to increase protein attachment to the bacterial cells. The effects of cations on protein attachment, bacterial adhesion, and hemagglutination were examined, and cation-binding components on both bacterial species were identified. The presence of cations, especially zinc, copper and cerium, increased attachment of human serum proteins to both bacterial species. In contrast, the presence of hydrophobic inhibitors or sugars had little effect. Protein attachment was reduced by heat treatment of the bacterial cells. Pretreatment of bacteria with human serum proteins inhibited adhesion of both species to buccal epithelial cells and hemagglutination. These effects were enhanced by the presence of zinc and copper during pretreatment. Using a chelating column, specific zinc- and copper-binding proteins were identified on the surfaces of both bacterial species.
Journal Article•
TL;DR: The pig-borne disease outbreaked recently in Sichuan province is supposed to be caused by Streptococcus suis type Ⅱ, which is highly virulent.
Abstract: Objective To isolate and identify the agent responsible for causing a high mortality in the infected people due to the recent zoonotic infection in Sichuan province.Methods Direct reverse transcript PCR and real time PCR were employed to detect the viral RNA of Nipah or Influenza A virus that has been suspected to be in the samples. The virus isolation was carried out with Vero cell and SPF Chicken embryos, and the cultures were subjected to PCR or hemagglutination(HA) test for Nipah or Influenza virus after each passage. Bacteria isolation were performed on blood nutrient agar from which the suspected colonies were then selected and tested by agglutination and PCR for Streptococcus suis type Ⅱ.Results The results showed that all the samples and their cultures were negative for influenza virus and Nipah virus in PCR and HA test, suggesting that the possibility of Nipah and Influenza A could be ruled out from the outbreak. However, three bacterial isolates from clinical samples were diagnosed as Streptococcus suis type Ⅱ based on Gram staining, biochemical character, agglutination and PCR test. PCR for virulent factor MRP (muramidase-released protein) and EF(extracellular factor) genes of Streptococcus suis type Ⅱ were all positive in the 3 isolates. Drug sensitivity tests revealed that all the isolates were highly sensitive to ampicillin, cephalothins Ⅵ, carbenicillin, compound sulfamethoxazole and cefuroxime.Conclusion The pig-borne disease outbreaked recently in Sichuan province is supposed to be caused by Streptococcus suis type Ⅱ, which is highly virulent.
Journal Article•
TL;DR: It is suggested that inactivated EDS antigen prepared by either heat or formalin treatment could successfully be used in avian diagnosis laboratories for routine flocks monitoring, EDS vaccine controlling and also surveillance of EDS vaccination program in poultry industry.
Abstract: Summary To prepare hemagglutination inhibition (HI) antigen the egg drop syndrome virus (EDSV) was propagated in 10-day-old embryonated duck eggs. The virus antigen was inactivated with two methods including heating (65°C) and adding 0.5% formaldehyde by considering low destroying effect on hemagglutination (HA) and HI titers. The EDSV HI titers of 310 sera and 100 yolks obtained from 23 chicken farms and specific pathogen free (SPF) chickens before and after EDS vaccination, in various ages were evaluated. Specificity, HA activity, stability and electron microscopic observations were similar in the prepared and standard antigens. The HI titers showed an equal sensitivity and specificity and, complete similarity (>99%) between the results obtained by prepared and standard antigens. Detection of antibody against EDSV using HI test showed slightly less sensitivity but more specificity compared with ELISA test. Upon the results it is suggested that inactivated EDS antigen prepared by either heat or formalin treatment could successfully be used in avian diagnosis laboratories for routine flocks monitoring, EDS vaccine controlling and also surveillance of EDS vaccination program in poultry industry.
Journal Article•
TL;DR: A method based on surface plasmon resonance (SPR) that detects the presence of the antigen-antibody complex without any labeling to rapidly quantitate anti-A/B IgG Ab levels is developed.
Abstract: The measurement of anti-blood group A/B (anti-A/B) IgG antibody levels is important for ABO unmatched-organ recipients because the effective removal of the antibodies improves their prognosis. Living-donor liver transplantation (LDLT) into ABO-unmatched patients tends to have a very poor outcome due to major complications such as intrahepatic bile duct complications and hepatic necrosis. Sustained bile duct complications are associated with high preoperative IgM type anti-A/B Ab titers, while patients with high preoperative IgG type anti-A/B Ab titers frequently develop sustained hepatic necrosis. There are several existing methods by which anti-A/B Ab levels can be measured, including the standard tube (TT) method, an enzyme-linked immunosorbent assay, and a flow cytometry method. Anti-A/B IgG Ab is difficult to identify by the TT method, which is the most popular method and is based on the detection of hemagglutination, because the major isotype that facilitates red cell agglutination is the pentameric IgM molecule. Therefore, we have developed a method based on surface plasmon resonance (SPR) that detects the presence of the antigen-antibody complex without any labeling. This method allows us to rapidly quantitate anti-A/B IgG Ab levels.