scholarly journals Large-Scale Survey of CampylobacterSpecies in Human Gastroenteritis by PCR and PCR–Enzyme-Linked Immunosorbent Assay

1999 ◽  
Vol 37 (12) ◽  
pp. 3860-3864 ◽  
Author(s):  
A. J. Lawson ◽  
J. M. J. Logan ◽  
G. L. O'neill ◽  
M. Desai ◽  
J. Stanley
Keyword(s):  
Immunosorbent Assay ◽  
Large Scale ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mixed Infections ◽  
Identification Algorithm ◽  
Detection And Identification ◽  
Significant Difference ◽  
Sporadic Cases ◽  
Low Incidence ◽  
Campylobacter Infection

A PCR-based study of the incidence of enteropathogenic campylobacter infection in humans was done on the basis of a detection and identification algorithm consisting of screening PCRs and species identification by PCR-enzyme-linked immunosorbent assay. This was applied to DNA extracted from 3,738 fecal samples from patients with sporadic cases of acute gastroenteritis, submitted by seven regional Public Health Laboratories in England and Wales over a 2-year period. The sending laboratories had cultured “Campylobacterspp.” from 464 samples. The PCR methodologies detected 492Campylobacter-positive samples, and the combination of culture and PCR yielded 543 Campylobacter-positive samples. There was identity (overlap) for 413 samples, but 79 PCR-positive samples were culture negative, and 51 culture-positive samples were PCR negative. While there was no statistically significant difference between PCR and culture in detection of C. jejuni-C. coli(PCR, 478 samples; culture, 461 samples), PCR provided unique data about mixed infections and non-C. jejuni and non- C. coli campylobacters. Mixed infections withC. jejuni and C. coli were found in 19 samples, and mixed infection with C. jejuni and C. upsaliensis was found in one sample; this was not apparent from culture. Eleven cases of gastroenteritis were attributed to C. upsaliensis by PCR, three cases were attributed to C. hyointestinalis, and one case was attributed to C. lari. This represents the highest incidence of C. hyointestinalis yet reported from human gastroenteritis, while the low incidence of C. larisuggests that it is less important in this context.

scholarly journals Development of a Recombinant Protein-Based Enzyme-Linked Immunosorbent Assay and Its Applications in Field Surveillance of Rodent Mice for Presence of Immunoglobulin G against Orientia tsutsugamushi

2003 ◽  
Vol 10 (3) ◽  
pp. 451-458 ◽  
Author(s):  
Yeau-Ching Wang ◽  
Ting-Yu Jian ◽  
Lih-Jeng Tarn ◽  
Yao-Wen Hung ◽  
Hai-Yuan Chao ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Immunosorbent Assay ◽  
Immunoglobulin G ◽  
Large Scale ◽  
Enzyme Linked Immunosorbent Assay ◽  
Orientia Tsutsugamushi ◽  
Suncus Murinus ◽  
Epitope Region ◽  
Increased Risk ◽  
Significant Difference

ABSTRACT A recombinant protein containing the immunodominant conserved epitope region of the 56-kDa outer membrane protein of the Karp strain of Orientia tsutsugamushi was purified to near homogeneity using recombinant DNA techniques. The purified protein was used to immunize rabbits and produced an antibody that could recognize different strains of O. tsutsugamushi, as demonstrated both by Western blotting and immunofluorescence assay. An enzyme-linked immunosorbent assay (ELISA) based on this recombinant protein was developed to detect antibody (immunoglobulin G [IgG]) against O. tsutsugamushi in mice captured in different districts of Taiwan during 2000 to 2001. A significant difference was found in the antibody seroprevalence rates of Suncus murinus mice captured in different districts of Taiwan (χ2 4, 0.95 = 26.64; P < 0.05). Furthermore, a significant difference of IgG seropositivity rates was observed among different kinds of mice (χ2 5, 0.95 = 93.85; P < 0.05). Antibody seropositivity rates were higher in Bandicota indica (100%), Rattus flavipectus (96.17%), and Rattus losea (95.83%) than in Rattus norvegicus (86.05%) and Rattus mindanensis (83.67%) (χ2 diff, 5, 0.95 = 12.59, P < 0.05). The lowest antibody seropositivity rate (54.4%) was observed in Suncus murinus. Antibody seropositivity rates of mice from different districts differed significantly because of the significant difference in antibody seroprevalence rates for S. murinus. The results of this study indicated that the recombinant protein ELISA developed in this study could be used to conduct large-scale surveillance of rodent mice for the presence of antibody against O. tsutsugamushi. The high seroprevalence rates in rodent mice (except S. murinus) suggest that people residing in these districts are at increased risk of developing O. tsutsugamushi infection.

Mechanization of the enzyme-linked immunosorbent assay (ELISA) for large scale screening of sera

1977 ◽  
Vol 16 (4) ◽  
pp. 351-359 ◽  
Author(s):  
E.J. Ruitenberg ◽  
J.A. van Amstel ◽  
B.J.M. Brosi ◽  
P.A. Steerenberg
Keyword(s):  
Immunosorbent Assay ◽  
Large Scale ◽  
Enzyme Linked Immunosorbent Assay ◽  
Large Scale Screening

scholarly journals DIFFERENCES OF SALIVA LEPTIN LEVELS IN CHILDREN WITH CHRONIC KIDNEY DISEASE ON HEMODIALYSIS AND HEALTHY CHILDREN (STUDY IN CHILDREN WITH GINGIVITIS)

2018 ◽  
Vol 11 (4) ◽  
pp. 192
Author(s):  
Berthauli Esther Nurmaida ◽  
Heriandi Sutadi ◽  
Sarworini B Budiardjo ◽  
Eka Laksmi Hidayati
Keyword(s):  
Chronic Kidney Disease ◽  
Kidney Disease ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Children ◽  
Significant Difference ◽  
The Difference

 Objectives: Analyze the difference of salivary leptin in between healthy children with gingivitis and hemodialysis (HD) children with gingivitis.Methods: A total of 20 children, ages 11–16-year-old with gingivitis, were chosen as subjects; 10 were on HD and 10 were healthy children. The level of salivary leptin was measured using the enzyme-linked immunosorbent assay method.Results: The results showed a significant difference of salivary leptin levels between the children on HD (61,300 ± 4151 pg/ml) and the healthy children (57,200 ± 3173 pg/ml).Conclusions: There is a significant difference in the salivary leptin levels in children on HD with gingivitis and healthy children with gingivitis.

Evaluation of immunoglobulin G4 subclass antibody in a peptide-based enzyme-linked immunosorbent assay for the serodiagnosis of human fascioliasis

Parasitology ◽  
2007 ◽  
Vol 134 (14) ◽  
pp. 2021-2026 ◽  
Author(s):  
C. TANTRAWATPAN ◽  
W. MALEEWONG ◽  
C. WONGKHAM ◽  
S. WONGKHAM ◽  
P. M. INTAPAN ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Large Scale ◽  
Laboratory Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Parasitic Infections ◽  
Immunoglobulin G4 ◽  
Predictive Values ◽  
Diagnostic Potential ◽  
Human Fascioliasis ◽  
Specific Igg4

SUMMARYTo improve the diagnosis of human fascioliasis caused byFasciola gigantica, we developed a peptide-based enzyme-linked immunosorbent assay (peptide-based ELISA) based on the detection of specific IgG4 subclass antibody. Two identified B-cell epitopes ofF. giganticacathepsin L1 were synthesized as single synthetic peptides, acetyl-DKIDWRESGYVTELKDQGNC-carboxamide (peptide L) and acetyl-DKIDWRESGYVTEVKDQGNC-carboxamide (peptide V), and their diagnostic potential was evaluated. The sera of 25 patients infected withF. gigantica, 212 patients with other parasitic infections, 32 cholangiocarcinoma patients and 57 healthy controls were analysed. The sensitivity, specificity, accuracy, and positive and negative predictive values of this assay were the same with both peptides at 100%, 99·7%, 99·7%, 96·2% and 100%, respectively. These highly sensitive and specific peptide-based ELISAs for the detection of specific IgG4 antibody could be useful for laboratory diagnosis of human fascioliasis in future large-scale surveys throughout Southeast Asia where this disease is prevalent.

Comparative evaluation of conventional staining method and enzyme linked immunosorbent assay kits for the detection of bovine cryptosporidiosis

2017 ◽  
Author(s):  
B. J. Thakre ◽  
J. B. Solanki ◽  
N. Kumar ◽  
A. Vargese
Keyword(s):  
Immunosorbent Assay ◽  
Staining Method ◽  
Dairy Calves ◽  
Occurrence Rate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Buffalo Calves ◽  
Significant Difference ◽  
Prevalence Difference ◽  
Cryptosporidium Parvum Oocysts ◽  
The Difference

Out of 428 faecal 178 (41.59%) animals were found positive for Cryptosporidium parvum oocysts in the bovine faeces in modified Ziehl-Neelsen (mZN) staining and/ or commercial plate and dipstick- enzyme linked immunosorbent assay (ELISA) kits (Cypress diagnostics, Langdorp, Belgium) with statistically non-significant difference in the occurrence rate in cattle and buffalo calves. Seasonal prevalence difference was statistically significant in cattle calves while non-significant in buffalo calves, respectively. The highest overall prevalence was recorded during rainy season (45.15%) and almost same per cent during winter and summer. Prevalence of 67.26, 37.11, 30 and 17.65% was recorded in calves aged below 1 month, 1-3 months, 4-8 months and 9-12 months, respectively (p>0.05). There was no significant difference in the age group prevalence between cattle and buffalo calves. Both sexes of bovines are equally susceptible to the cryptosporidiosis (p>0.05). The prevalence of cryptosporidiosis was 59.54 and 29.41% for diarrhoeic and non-diarrhoeic samples, respectively (p>0.05). Both diarrhoeic and non-diarrhoeic groups of calves aged between 1-30 days were equally susceptible to the infection of Cryptosporidium spp. Almost same per cent prevalence of the infection was observed in dairy calves reared in organized (40.76%) and unorganized farms (42.21%) and the difference was non-significant in both cattle and buffalo calves. Highest prevalence of cryptosporidiosis was found in HF cross calves. The tests failed to detect the oocysts in infected soil samples. There was highly significant difference in the prevalence of cryptosporidiosis estimated by plate/ dipstick-ELISA and mZN staining with highest sensitivity and specificity in plate-ELISA.

scholarly journals In Situ Immunoassay (ISIA) of Field Grapefruit Trees Inoculated with Mild Isolates of Citrus tristeza virus Indicates Mixed Infections with Severe Isolates

Plant Disease ◽  
2002 ◽  
Vol 86 (5) ◽  
pp. 458-461 ◽  
Author(s):  
Youjian Lin ◽  
Phyllis A. Rundell ◽  
Charles A. Powell
Keyword(s):  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
Citrus Tristeza Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mixed Infections ◽  
The Cross ◽  
Tristeza Virus ◽  
Mild Isolate ◽  
Citrus Tristeza

Ten grapefruit trees that had been inoculated with a mild isolate of Citrus tristeza virus (CTV) and maintained in the field for 18 years were found in a previous study to be declining and infected with severe isolates of CTV, or symptomless and infected with mild isolates of CTV, using enzyme-linked immunosorbent assay (ELISA). They were assayed with an in situ immunoassay (ISIA) procedure using monoclonal antibodies 17G11 (reacts with most Florida isolates of CTV) and MCA13 (reacts with severe, but not Florida mild isolates of CTV). All the grapefruit trees were 17G11 positive by ELISA and ISIA. The five trees that showed moderate decline symptoms were MCA13 positive by ELISA and ISIA. The five symptomless trees were MCA13 negative by ELISA. However, four of the five symptomless trees were MCA13 positive by ISIA, which showed that ISIA with MCA13 had greater sensitivity in detecting severe CTV isolates than ELISA. These results suggested that the cross-protected grapefruit trees, regardless of symptoms, were infected with both mild and severe isolates of CTV.

Seroepidemiology of Toxoplasma gondii in pet dogs and cats in Beijing, China

2008 ◽  
Vol 53 (3) ◽  
Author(s):  
Jinhai Yu ◽  
Jun Ding ◽  
Zhaofei Xia ◽  
Degui Lin ◽  
Yili Li ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Immunosorbent Assay ◽  
Age Groups ◽  
Latex Agglutination ◽  
Agglutination Test ◽  
Latex Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Pet Dogs ◽  
Significant Difference ◽  
Beijing China

AbstractSera from 534 pet dogs and 335 pet cats from Beijing (China) were tested for anti-Toxoplasma gondii antibodies using an enzyme-linked immunosorbent assay or the latex agglutination test. The seropositivity by year, season, sex and age was analysed. Overall, 128 dogs (24.0%) and 50 cats (14.9%) had antibodies to T. gondii. When analysed by season, the highest seroprevalence was found in spring for dogs (31.3%) and cats (25.1%), and the differences in seroprevalence by season was statistically significant in cats (P<0.01) but not in dogs. The seroprevalence in male dogs (23.7%) and cats (15.1%) were slightly higher than their female counterparts (18.0% in dogs and 12.3% in cats). There was no obvious pattern of seropositivity or significant difference in different age groups in dogs or cats; nonetheless, a high proportion of dogs at 4 years of age were positive to T. gondii (31.8%) while cats with relatively high seropositivity rates were at 1 or 3.4 years of age (13.14%).

scholarly journals Evaluation of Cross Protective Efficacy of Commercial Vaccines against Mannheimia haemolytica in Mice

2019 ◽  
Vol 43 (1) ◽  
pp. 165-170
Author(s):  
Waffa A. Ahmed
Keyword(s):  
Immunosorbent Assay ◽  
Protective Efficacy ◽  
Challenge Test ◽  
Cross Protection ◽  
Mannheimia Haemolytica ◽  
Economic Losses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Significant Difference ◽  
Protection Efficacy

Mannheimia haemolytica together with Pasteurella multocida represents as a major bacterial causative agent of cattle, sheep and goats respiratory diseases and its one of the most important causes for economic losses to these animals .Commercially available vaccines were used to prevent infections caused by P. multocida and M. haemolytica. Thus, the aim of the present study was to evaluate the cross protection efficacy of two vaccines to protect mice against M.haemolytica, studying humeral immunity, using Enzyme-Linked Immunosorbent Assay. Forty five mice were divided into three equal groups, group one and two were inoculated subcutaneously  4μl\JOVAPAST® and 1μl of Al-kindy vaccines respectively, while the third group was with 0.5 ml sub cutaneous PBS. LD50for M.haemolytica was estimated as 2× 106 cfu \ml and challenge test was conducted by dropping 0.05 ml 2× 106 cfu \ml intranasally after three weeks of immunization for the three groups. The results of Enzyme-Linked Immunosorbent Assay, showed significant increase of antibody titters at (P<0.01) in (group 1 and 2) after first and second weeks post immunization, in comparison with control group. Also, the re-isolation of M.haemolytica from lungs tissue of all groups after challenged were positive with significant difference between control and immunized group, control group was 4× 108 cfu ∕ml which was higher than immunized group one and group two,which were 2.5×104 cfu∕ml and 3,5×105 cfu∕ml respectively after 24 hour of vaccine. In conclusion, the two commercial vaccines showed good cross protection efficacy against M. haemolytica, but JOVAPAST® vaccine showed higher efficacy than Alkindy vaccine, as that it contain  two  heterologous  killed strains and providing the basis for production a vaccine from the two  pathogen of local strains. 

156 THE USE OF THE DG29™ ENZYME-LINKED IMMUNOSORBENT ASSAY KIT TO PREDICT PREGNANCY PRIOR TO EMBRYO TRANSFER IN LACTATING HOLSTEIN COWS

2013 ◽  
Vol 25 (1) ◽  
pp. 226
Author(s):  
Y. Nakamura ◽  
A. Ideta ◽  
A. Shirasawa ◽  
K. Hayama ◽  
S. Sakai ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Statistical Significance ◽  
Visual Observation ◽  
Holstein Cows ◽  
Enzyme Linked Immunosorbent Assay ◽  
Animal Reproduction ◽  
Array Transducer ◽  
Significant Difference ◽  
Chi Squared ◽  
Student’S T

Evaluation of postpartum fertility in cows is important for the efficient management of reproduction. DG29™ enzyme-linked immunosorbent assay (ELISA) kit (Conception, Animal Reproduction Technologies, Canada) measures the level of pregnancy–related glycoproteins in blood that are linked to pregnancy in the bovine species. The proteins are known to persist in the postpartum period. Here, we investigated whether the postpartum fertility in Holstein dairy cows can be evaluated through the use of the DG29 kit. We confirmed that genital organs of lactating Holstein cows (n = 119, from Days 56 to 688 postpartum) were normal by a 5.0/7.5-MHz linear array transducer (Tringa, Pie Medical Equipment B.V., Maastricht, The Netherlands), then a progesterone releasing intravaginal device (PRID; CEVA Sante Animale, Libourne, France) was inserted (Day 0) and maintained for 9 days. On Day 7, PGF2α was administered (2 mL Dalmazine, Kyoritsu Seiyaku, Tokyo, Japan). Blood samples were collected from the tail vein or artery into vacuum tubes at the time of PRID insertion. Serum was separated and stored at –30°C until the ELISA was performed. Oestrus (Day 0) was detected by visual observation. Fresh embryos recovered from Japanese Black cows were transferred to 119 recipient cows in various parities (primiparous = 70, biparous = 27, and multiparous = 22) on Days 6 to 8 of oestrous cycle. Pregnancy was diagnosed between Days 40 to 60 by transrectal ultrasonography. The statistical significance of any differences between various parities was assessed by chi-squared and Student’s t-tests. The pregnancy rate was higher for primiparous cows than for biparous and multiparous cows (64.3, 55.6, and 54.5%, respectively), while concentrations of the pregnancy-related glycoproteins in primiparous cows (135.0 ± 29.8 pg mL–1) were significantly lower than those of biparous (389.4 ± 175.9 pg mL–1) and multiparous cows (399.2 ± 203.1 pg mL–1, mean ± SEM; P < 0.05). In primiparous and multiparous cows, the concentrations of pregnancy-related glycoproteins were significantly lower in pregnant cows compared with nonpregnant cows (primiparous: 81.1 ± 29.9 v. 232.6 ± 59.8 pg mL–1; P < 0.05; multiparous: 20.8 ± 16.2 v. 853.4 ± 411.5 pg mL–1; P < 0.05). However, there was no significant difference between pregnant and nonpregnant biparous cows. In conclusion, the DG29 kit may be useful for the prediction of postpartum fertility in lactating Holstein cows. Further studies are needed to test the validity of this observation by using a greater number of various parties’ cows.

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