scholarly journals Evaluation of “Indigenous Absorbed ELISA Kit” for the Estimation of Seroprevalence of Mycobacterium avium Subspecies paratuberculosis Antibodies in Human Beings in North India

2011 ◽  
Vol 2011 ◽  
pp. 1-5 ◽  
Author(s):  
A. V. Singh ◽  
S. V. Singh ◽  
D. K. Verma ◽  
R. Yadav ◽  
P. K. Singh ◽  
...  

In present pilot study aimed to estimate, presence of Mycobacterium avium subspecies paratuberculosis (MAP) antibodies in the human serum samples originating from North India using “Indigenous absorbed ELISA kit” (ELISA kit). The phase I, “ELISA kit” was optimized using protoplasmic antigen from native isolate of MAP “Indian Bison type” recovered from the biopsies of Crohn's disease patients. The phase II, sensitivity and specificity of the kit were estimated as 40.0 and 83.3%, respectively, when evaluated in 40 human serum samples (5 Crohn's disease and 22 ulcerative colitis patients and 13 healthy human subjects) with defined MAP status with respect to stool culture. Seroprevalence of MAP antibodies was higher in CD patients (80.0%) as compared to ulcerative colitis patients (4.5%) and normal human subjects (15.3%). The phase III, seroprevalence of MAP antibodies was estimated as 23.4%, on the basis of the screening of 452 human serum samples (without history) from different geographical regions of North India. Region-wise, 34.0, 33.3, 32.8, 25.0, 23.0, 17.7, and 12.5% samples were positive from the states of Punjab, Uttarakhand, New Delhi, Himachal Pradesh, Haryana, Uttar Pradesh and Jammu and Kashmir, respectively. Study reported moderately higher presence of MAP antibodies in human population, which necessitates programs to reduce the bioburden of MAP in the environment and in animal population.

Author(s):  
Sara Notararigo ◽  
Manuel Martín-Pastor ◽  
Juan E. Viñuela Roldán ◽  
Adriano Quiroga ◽  
J. Enrique Dominguez-Munoz ◽  
...  

Abstract Inflammatory bowel disease is a multifactorial etiology, associated with environmental factors that can trigger both debut and relapses. A high level of tumor necrosis factor-α in the gut is the main consequence of immune system imbalance. The aim of treatment is to restore gut homeostasis. In this study, fresh blood and serum samples were used to identify biomarkers and to discriminate between Crohn’s disease and ulcerative colitis patients under remission treated with anti-TNF. Metabolomics based on Nuclear Magnetic Resonance spectroscopy (NMR) was used to detect unique biomarkers for each class of patients. Blood T lymphocyte repertories were characterized, as well as cytokine and transcription factor profiling, to complement the metabolomics data. Higher levels of homoserine-methionine and isobutyrate were identified as biomarkers of Crohn’s disease with ileocolic localization. For ulcerative colitis, lower levels of creatine-creatinine, proline, and tryptophan were found that reflect a deficit in the absorption of essential amino acids in the gut. T lymphocyte phenotyping and its functional profiling revealed that the overall inflammation was lower in Crohn’s disease patients than in those with ulcerative colitis. These results demonstrated that NMR metabolomics could be introduced as a high-throughput evaluation method in routine clinical practice to stratify both types of patients related to their pathology. Key messages NMR metabolomics is a non-invasive tool that could be implemented in the normal clinical practice for IBD to assess beneficial effect of the treatment. NMR metabolomics is a useful tool for precision medicine, in order to sew a specific treatment to a specific group of patients. Finding predictors of response to IFX would be desirable to select patients affected by IBD. Immunological status of inflammations correlates with NMR metabolomics biomarkers.


2020 ◽  
Vol 14 (Supplement_1) ◽  
pp. S304-S304
Author(s):  
A HOLMER ◽  
B Boland ◽  
S Singh ◽  
H Le ◽  
J Neill ◽  
...  

Abstract Background The endoscopic healing index (EHI, Monitr, Prometheus Biosciences, San Diego, CA) is a serum-based biomarker panel available for identifying mucosal inflammation in Crohn’s disease.[1] We aimed to study its performance for identifying mucosal inflammation in ulcerative colitis. Methods EHI was analysed on serum samples paired with endoscopies from adult patients (≥18 years) participating in a prospective biobank (June 2014 to December 2017). Area under receiver operating characteristic curves (AUROC) were used to assess the accuracy of EHI for endoscopic improvement (EI; Mayo endoscopic sub-score [MES] 0–1) and endoscopic remission (ER; MES 0). Sensitivity for EHI was calculated using a cut-off previously identified for Crohn’s disease which optimised performance for ruling out endoscopic activity (20 points). Alternative cut-offs were explored. Results A total of 114 patients were included, with an overall prevalence of 56% and 44% for EI and ER. The AUROC was 0.79 (95% CI 0.70–0.87) for EI and 0.70 (95% CI 0.61–0.80) for ER. A cut-off of 20 points had a sensitivity of 94% (95% CI 83–99%) for ruling out moderate to severe (MES 2–3) endoscopic activity, and a sensitivity of 84% (95% CI 72–92%) for ruling out mild to severe (MES 1–3) endoscopic activity. A cut off of 40 points or higher had > 90% specificity for ruling in moderate to severe (MES 2–3) or mild to severe (MES 1–3) endoscopic activity. (Table 1) Conclusion EHI has favourable accuracy in identifying the presence of mucosal inflammation in patients with ulcerative colitis. Although it was not developed and validated for ulcerative colitis, further validation is warranted. Reference


2005 ◽  
Vol 134 (1) ◽  
pp. 49-56 ◽  
Author(s):  
P. H. JONES ◽  
T. B. FARVER ◽  
B. BEAMAN ◽  
B. ÇETINKAYA ◽  
K. L. MORGAN

SUMMARYMycobacterium aviumsubspeciesparatuberculosis(Map), the cause of ruminant paratuberculosis, has been proposed as the causative agent of Crohn's disease. The objective of this study was to determine whether exposure to clinical cases of bovine paratuberculosis was a risk factor for Crohn's disease. A questionnaire was sent to dairy farmers living on premises where the occurrence or absence of clinical cases of bovine paratuberculosis had previously been determined. The prevalence of Crohn's disease was found to be similar to that reported in other studies in the United Kingdom and showed no association with bovine paratuberculosis. There was, however, a univariate association with geographical region. Ulcerative colitis showed univariate associations with age, frequency of contact with cattle and with smoking. The results do not support the hypothesis thatMapplays a causative role in the aetiology of Crohn's disease.


2020 ◽  
Vol 8 (12) ◽  
pp. 2054
Author(s):  
J. Todd Kuenstner ◽  
Raghava Potula ◽  
Tim J. Bull ◽  
Irene R. Grant ◽  
Antonio Foddai ◽  
...  

Mycobacterium avium subspecies paratuberculosis (MAP) has long been suspected to be involved in the etiology of Crohn’s disease (CD). An obligate intracellular pathogen, MAP persists and influences host macrophages. The primary goals of this study were to test new rapid culture methods for MAP in human subjects and to assess the degree of viable culturable MAP bacteremia in CD patients compared to controls. A secondary goal was to compare the efficacy of three culture methods plus a phage assay and four antibody assays performed in separate laboratories, to detect MAP from the parallel samples. Culture and serological MAP testing was performed blind on whole blood samples obtained from 201 subjects including 61 CD patients (two of the patients with CD had concurrent ulcerative colitis (UC)) and 140 non-CD controls (14 patients in this group had UC only). Viable MAP bacteremia was detected in a significant number of study subjects across all groups. This included Pozzato culture (124/201 or 62% of all subjects, 35/61 or 57% of CD patients), Phage assay (113/201 or 56% of all subjects, 28/61 or 46% of CD patients), TiKa culture (64/201 or 32% of all subjects, 22/61 or 36% of CD patients) and MGIT culture (36/201 or 18% of all subjects, 15/61 or 25% of CD patients). A link between MAP detection and CD was observed with MGIT culture and one of the antibody methods (Hsp65) confirming previous studies. Other detection methods showed no association between any of the groups tested. Nine subjects with a positive Phage assay (4/9) or MAP culture (5/9) were again positive with the Phage assay one year later. This study highlights viable MAP bacteremia is widespread in the study population including CD patients, those with other autoimmune conditions and asymptomatic healthy subjects.


2019 ◽  
Vol 37 (15_suppl) ◽  
pp. 2582-2582
Author(s):  
Shilpa Grover ◽  
Amitabh Srivastava ◽  
Sonia Friedman ◽  
Elizabeth Iannotti Buchbinder ◽  
F. Stephen Hodi ◽  
...  

2582 Background: Colitis is a frequent toxicity of ICI therapy but there is paucity of data on risk factors. Specific serological markers and genetic polymorphisms have been associated with inflammatory bowel disease (IBD) (ulcerative colitis, Crohn’s disease). However, the prevalence of these markers in pts with ICI colitis is unknown. We performed a pilot study to determine the prevalence of IBD-associated genetic and serologic biomarkers in pts with ICI colitis. Methods: Cancer pts with histologically confirmed ICI enteritis/colitis and no history of IBD underwent commercial IBD panel testing. The panel included 4 genetic markers ( ATG16L1, NXK2–3, ECM1, STAT3), 8 serological markers (anti-A4-Fla2, anti-A4-FlaX, anti-CBir1, anti-OmpC, ASCAIgA, ASCA-IgG, pANCA, ANCA), and 5 inflammatory markers (vascular endothelial growth factor [VEGF], intracellular adhesion molecule 1[ICAM-1], vascular cell adhesion molecule 1 [VCAM-1], C-reactive protein, serum amyloid A [SAA]). Clinical testing on serum samples was performed by Prometheus Laboratories (San Diego, CA). Results: Of 15 cancer pts with biopsy confirmed ICI colitis, 10 (67%) were homozygous for 1 or more of 4 genetic markers. The remaining 5 pts were all heterozygous for two or more of the genetic markers. One or more serologic markers associated with IBD were elevated in 7/15 (47%) pts. Serum reactivity was noted for ASCA-IgA (1/15, 7%), ASCA-IgG (1/15, 7%), anti-OmpC (3/15, 20%), anti-CBR IgG (2/15, 13%), anti-A4-FlaX (1/15, 7%), and ANCA (2/15, 13%). One or more inflammatory markers were elevated in 13/15 (88%) pts. Elevations in VEGF, VCAM-1, ICAM-1, and SAA were noted in 2 (13%), 8 (53%), 8 (53%), and 11 (73%) pts, respectively. Only 6 (40%) pts had elevations in CRP levels despite the presence of active inflammation on biopsy. The IBD panel was reported as being consistent with Crohn’s disease in 2 pts, ulcerative colitis in 1 pt and inconclusive for type but consistent with IBD in 1 pt. Conclusions: In this pilot study, all patients with ICI colitis, were either homozygous or heterozygous for two or more high risk IBD alleles. If validated, such testing may prospectively identify pts at risk for developing ICI colitis.


Gut ◽  
1998 ◽  
Vol 43 (3) ◽  
pp. 375-382 ◽  
Author(s):  
W Reinisch ◽  
K-H Heider ◽  
G Oberhuber ◽  
C Dejaco ◽  
M Müllner ◽  
...  

Background—Increased expression of CD44v6 on colonic crypt epithelial cells in ulcerative colitis has been suggested as a diagnostic tool to distinguish ulcerative colitis from colonic Crohn’s disease.Aims—To investigate colonic CD44v6 expression and serum concentrations of soluble CD44v6 (sCD44v6) in patients with ulcerative colitis and Crohn’s disease.Methods—Colonic biopsy samples were obtained from 16 patients with ulcerative colitis, 13 with ileocolonic Crohn’s disease, and 10 undergoing polypectomy. Serum samples were obtained from 15 patients with active ulcerative colitis, 20 with active Crohn’s disease, and 20 healthy donors. Colonic CD44v6 expression was evaluated immunohistochemically by monoclonal antibody 2F10 and the higher affinity monoclonal antibody VFF18. Serum sCD44v6 concentrations were measured by ELISA.Results—2F10 stained colonic epithelium of inflamed ulcerative colitis and Crohn’s disease samples in 80% and 40% of cases, respectively, and VFF18 in 95% and 87%, respectively. Both monoclonal antibodies displayed a sensitivity and specificity of 60% and 87% to differentiate ulcerative colitis from colonic Crohn’s disease. Serum concentrations of sCD44v6 were lower in patients with ulcerative colitis (median 153 ng/ml; interquartile range (IQR) 122–211) compared with Crohn’s disease (219; IQR 180–243) and healthy donors (221; IQR 197–241 (p=0.002)). Its sensitivity and specificity to discriminate ulcerative colitis from Crohn’s disease was 75% and 71%, respectively.Conclusion—Colonic CD44v6 and serum sCD44v6 concentrations do not facilitate reliable differential diagnosis between ulcerative colitis and Crohn’s disease.


2020 ◽  
Vol 14 (Supplement_1) ◽  
pp. S138-S139
Author(s):  
F Fart ◽  
S Salihovic ◽  
A McGlinchey ◽  
M Orešič ◽  
J Halfvarson ◽  
...  

Abstract Background Environmental factors have been implicated in the pathogenesis of inflammatory bowel disease (IBD), particularly late onset disease. Per- and polyfluoroalkyl substances (PFAS) are man-made chemicals with a long biological half-life that have been extensively used since the 1950s and have been proposed to interfere with the bile acid synthesis. Therefore, to investigate if late onset IBD correlates with higher PFAS levels, we measured serum levels of PFAS and bile acids in patients diagnosed with IBD later in life. Methods Serum samples were collected from patients diagnosed with ulcerative colitis (n = 20) and Crohn’s disease (n = 20) at the age of ≥55 years. Blood donors (n = 20) were used as healthy controls and were matched by gender and age. The levels of PFAS and bile acids were assessed by ultra performance liquid chromatography coupled to a triple quadrupole mass spectrometer. Results The total amount of PFAS was significantly higher in patients with ulcerative colitis compared with healthy controls (p = 0.021) or patients with Crohn’s disease (p = 0.015). No difference was found in total PFAS levels between Crohn’s disease patients and healthy controls (p = 0.841). Seven out of 30 bile acids correlated to the total PFAS level. Conclusion Our results demonstrate that PFAS levels are increased in patients with late-onset of ulcerative colitis compared with Crohn’s disease patients and healthy controls. This finding indicates that PFAS might represent an environmental risk factor for ulcerative colitis. However, additional studies assessing the functional consequences of increased PFAS in late-onset ulcerative colitis are required to confirm this hypothesis.


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