High prevalence of Mycobacterium avium subspecies paratuberculosis (‘Indian bison type’) in animal attendants suffering from gastrointestinal complaints who work with goat herds endemic for Johne's disease in India

Objectives

In this study we aimed to estimate the prevalence of Mycobacterium avium subspecies paratuberculosis (MAP) in animal attendants who were chronic colitis patients or who had inflammatory bowel disease and were suspected for Crohn's disease; these animal attendants worked with goat herds endemic for Johne's disease. Microscopic examination and culture tests were used. For comparison purposes a group of healthy human subjects (not suffering with colitis) was also screened.

Methods

Stool samples obtained from 98 human subjects (58 animal attendants suspected for Crohn's disease and 40 healthy humans) were screened for the presence of MAP by microscopic examination and culture. Of the 58 animal attendants screened, 38 had abdominal pain, 29 had suffered episodes of diarrhea, 39 had experienced weight loss, 27 had fever, and 32 had a history of raw milk consumption. Animal attendants had had contact of variable duration with goat herds endemic for Johne's disease (1–5, 6–10, 11–15, and >15 years). Forty stool samples from healthy humans with no symptoms/history of contact with animals were also screened. IS900 PCR and IS1311 PCR restriction endonuclease analysis were used to characterize and genotype the MAP colonies.

Results

MAP was recovered from 34 of the 98 human subject stool samples (34.7%). Of the 98 samples, 16.3% (n = 16) were acid-fast. None of the 40 healthy human subjects were positive for MAP by microscopy, but five (12.5%) were positive for MAP by culture. Of the 58 animal attendants, 16 (27.6%) were positive by microscopy and 29 (50%) were positive by culture. MAP were recovered from 68.4% of animal attendants with abdominal pain, 72.4% of those with diarrhea, 71.8% of those with weight loss, 44.4% of those with fever, and 46.9% of those who had a history of raw milk consumption. Of the 29 culture-positive animal attendants, 48.3% had worked for >15 years, 27.6% for 11–15 years, 20.7% for 6–10 years, and 3.4% for 1–5 years with goat herds endemic for Johne's disease. Of the 34 culture isolates, 28 (82.4%) showed good quality DNA on agarose gel and were positive by IS900 PCR. Of the 28 IS900-positive DNA samples, 23 (82.1%) were genotyped as ‘Indian bison type’ and five (17.9%) as ‘cattle type’.

Conclusions

The prevalence of MAP was higher in attendants suffering from gastrointestinal problems who worked with goat herds endemic for Johne's disease, than in humans with no history of contact with animals. The risk of developing gastrointestinal problems with clinical symptoms indistinguishable from inflammatory bowel disease was higher in humans who were in contact with goat herds endemic for Johne's disease as compared to healthy humans, and the risk was correlated with the duration of association with the endemic goat herds.     

Genotype profiles of Mycobacterium avium subspecies paratuberculosis recovered from suspected and Crohn's disease patients in India

Present study aimed to genotype Mycobacterium avium subspecies paratuberculosis (MAP) recovered from suspected and Crohn' s disease patients. A total of 32 MAP and DNA (directly from clinical samples) recovered from human origin were genotyped using IS 1311 PCR-REA. Isolates were cultured from stool, biopsies and blood clots of Crohn's disease patients, and stool samples of suspected (animal attendants, lab workers etc). Of the 32 MAP isolates belonging to 28 human beings, majority (84.3%) were genotyped as 'Bison type', while 21.7% were of 'cattle' and none was 'sheep' genotype. Study first time reports distribution of 'Cattle' and 'Bison type' 'genotypes in suspected and Crohn's patients on pilot scale in India. 'Bison type' genotype was predominant in the surveyed human population.     

Is Crohn's disease caused by a mycobacterium? Comparisons with leprosy,tuberculosis, and Johne's disease

Although Crohn's disease is considered to be autoimmune in origin, there is increasing evidence that it may have an infectious cause. The most plausible candidate is Mycobacterium avium subspecies paratuberculosis (MAP). Intriguingly, Koch's postulates may have been fulfilled for MAP and Crohn's disease, even though they still have not been met for Mycobacterium leprae and leprosy. In animals MAP causes Johne's disease, a chronic wasting intestinal diarrhoeal disease evocative of Crohn's disease. Johne's disease occurs in wild and domesticated animals, including dairy herds. Viable MAP is found in human and cow milk, and is not reliably killed by standard pasteurisation. MAP is ubiquitous in the environment including in potable water. Since cell-wall-deficient MAP usually cannot be identified by Ziehl-Neelsen staining, identification of MAP in human beings requires culture or detection of MAP DNA or RNA. If infectious in origin, Crohn's disease should be curable with appropriate antibiotics. Many studies that argue against a causative role for MAP in Crohn's disease have used antibiotics that are inactive against MAP. However, trials that include macrolide antibiotics indicate that a cure for Crohn's disease is possible. The necessary length of therapy remains to be determined. Mycobacterial diseases have protean clinical manifestations, as does Crohn's disease. The necessity of stratifying Crohn's disease into two clinical manifestations (perforating and non-perforating) when interpreting the results of antibiotic therapy is discussed. Rational studies to evaluate appropriate therapies to cure Crohn's disease are proposed.     

Lack of support for a common etiology in Johne's disease of animals and Crohn's disease in humans.

The superficial similarity of Johne's disease to Crohn's disease led to the hypothesis that, like the former. Crohn's disease was caused by Mycobacterium paratuberculosis. Detailed pathologic comparisons, however, reveal little similarity between these two entities, including the lack of important extraintestinal manifestations. Attempts to recover M. paratuberculosis by culture have only rarely succeeded and the significance of spheroplasts that appear more frequently on culture is seriously in question. Five immunocytochemistry studies have failed to find mycobacterial antigens in diseased tissues and the five most recent polymerase chain reaction (PCR) attempts to find genomic evidence of M. paratuberculosis were uniformly negative. Numerous serologic studies failed to demonstrate antibody to M. paratuberculosis and attempts to show cell-mediated immunity were also unrewarding. Inoculation of numerous experimental animals with Crohn's disease tissue has failed to induce Johne's disease, and inoculation of various animal species with M. paratuberculosis has equally failed to result in Crohn's disease. Controlled studies of the treatment of Crohn's disease with antimycobacterial agents have generally resulted in no improvement, and most studies that have shown a positive response are either uncontrolled or include broad-spectrum antibiotics that may be acting on pathogens other than mycobacteria. Finally, although Johne's disease is common in farm animals, and infected animals shed M. paratuberculosis in large numbers, no record of zoonotic transmission has been recorded.     

Ulcerative colitis and Crohn's disease: is Mycobacterium avium subspecies paratuberculosis the common villain?

Mycobacterium avium, subspecies paratuberculosis (MAP) causes a chronic disease of the intestines in dairy cows and a wide range of other animals, including nonhuman primates, called Johne's ("Yo-knee's") disease. MAP has been consistently identified by a variety of techniques in humans with Crohn's disease. The research investigating the presence of MAP in patients with Crohn's disease has often identified MAP in the "negative" ulcerative colitis controls as well, suggesting that ulcerative colitis is also caused by MAP. Like other infectious diseases, dose, route of infection, age, sex and genes influence whether an individual infected with MAP develops ulcerative colitis or Crohn's disease. The apparently opposite role of smoking, increasing the risk of Crohn's disease while decreasing the risk of ulcerative colitis, is explained by a more careful review of the literature that reveals smoking causes an increase in both diseases but switches the phenotype from ulcerative colitis to Crohn's disease. MAP as the sole etiologic agent of both ulcerative colitis and Crohn's disease explains their common epidemiology, geographic distribution and familial and sporadic clusters, providing a unified hypothesis for the prevention and cure of the no longer "idiopathic" inflammatory bowel diseases.     

Mycobacterium avium subspecies paratuberculosis and Crohn's disease: a systematic review and meta-analysis

This systematic review assesses the evidence for an association between Mycobacterium avium subspecies paratuberculosis (MAP) and Crohn's disease. We analysed 28 case-control studies comparing MAP in patients with Crohn's disease with individuals free of inflammatory bowel disease (IBD) or patients with ulcerative colitis. Compared with individuals free of IBD, the pooled odds ratio (OR) from studies using PCR in tissue samples was 7.01 (95% CI 3.95-12.4) and was 1.72 (1.02-2.90) in studies using ELISA in serum. ORs were similar for comparisons with ulcerative colitis patients (PCR, 4.13 [1.57-10.9]; ELISA, 1.88 [1.26-2.81]). The association of MAP with Crohn's disease seems to be specific, but its role in the aetiology of Crohn's disease remains to be defined.     

Detection and Isolation of Mycobacterium avium subspecies paratuberculosis from intestinal mucosal biopsies of patients with and without Crohn's disease in Sardinia

OBJECTIVES: Sardinia is an island community of 1.6 million people. There are also about 3.5 million sheep and one hundred thousand cattle in which Johne's disease and Mycobacterium avium subspecies paratuberculosis infection are endemic. The present study was designed to determine what proportion of people in Sardinia attending for ileocolonoscopy with or without Crohn's disease were infected with this pathogen. METHODS: Mycobacterium avium subspecies paratuberculosis was detected by IS900 PCR on DNA extracts of fresh intestinal mucosal biopsies as well as by isolation in culture using supplemented MGIT media followed by PCR with amplicon sequencing. RESULTS: Twenty five patients (83.3%) with Crohn's disease and 3 control patients (10.3%) were IS900 PCR positive (p = 0.000001; Odds ratio 43.3). Mycobacterium avium subspecies paratuberculosis grew in cultures from 19 Crohn's patients (63.3%) and from 3 control patients (10.3%) (p = 0.00001; Odds ratio 14.9). All patients positive by culture had previously been positive by PCR. Mycobacterium avium subspecies paratuberculosis first appeared in the liquid cultures in a Ziehl Neelsen (ZN) staining negative form and partially reverted through a rhodamine-auramine positive staining form to the classical ZN positive form. This resulted in a stable mixed culture of all 3 forms illustrating the phenotypic versatility of these complex chronic enteric pathogens. CONCLUSIONS: Mycobacterium avium subspecies paratuberculosis was detected in the majority of Sardinian Crohn's disease patients. The finding of the organism colonizing a proportion of people without Crohn's disease is consistent with what occurs in other conditions caused by a primary bacterial pathogen in susceptible hosts.     

Bile acid studies in patients with Crohn's colitis.

Bile acid studies were performed in patients with Crohn's disease, radiologically confined to the colon. The bile acid pool size of 10 patients with isolated Crohn's colitis was significantly lower than that of 10 normal control subjects (P less than 0.001) and of 10 ulcerative colitis patients (P less than 0.005). Measurements of 14C-excretion in breath and in 24 hours stool collections after the administration of 5 muCi 14C-glycocholate showed a normal 14C-excretion in breath and usually a markedly increased loss of 14C in the stool (greater than 7% of the dose). The simultaneous administration of 5 muCi 3H-polyethylene glycol MW 4000 (3H-PEG) as a marker indicated that the 14C/3H ratio in the patients with Crohn's colitis was significantly greater than in a control series of patients with diarrhoea not due to bile acid malabsorption. Studies on the composition of duodenal bile showed a significantly decreased concentration of deoxycholic acid in duodenal bile. These observations suggest bile acid malabsorption in patients with Crohn's disease apparently confined to the colon.     

Mycobacterial aetiology of Crohn's disease: serologic study using common mycobacterial antigens and a species-specific glycolipid antigen from Mycobacterium paratuberculosis.

Crohn's disease is a granulomatous form of enteritis superficially similar to Johne's disease (paratuberculosis) of ruminants. Recently, a Mycobacterium sp closely related to Mycobacterium paratuberculosis was cultured from tissues of patients with Crohn's disease suggesting that M paratuberculosis may be the aetiologic agent in some cases. In addition, greater seroreactivity to M paratuberculosis has been reported in patients with Crohn's disease. In the present study, we have evaluated the serum antibody response to disrupted M paratuberculosis using ELISA and serum specimens from 33 people with Crohn's disease, 21 with ulcerative colitis, and 12 non-inflammatory bowel disease controls. We failed to find a consistent IgG, IgM, or IgA antibody response to Mycobacterium paratuberculosis. The results indicate that, as in bovine paratuberculosis, serum seroreactivity is not a reliable tool for examining the relationship between human intestinal disease and mycobacteria.     

Mycobacterium paratuberculosis DNA not detected in Crohn's disease tissue by fluorescent polymerase chain reaction.

The role of mycobacteria in the aetiology of Crohn's disease has been a contentious subject for many years. Mycobacterium paratuberculosis is known to cause a chronic granulomatous enteritis in animals (Johne's disease) and has been implicated as a possible infectious cause of Crohn's disease. However this fastidious organism is only rarely detected by conventional microbiological techniques. This study used oligonucleotide primers to the species-specific M paratuberculosis IS900 DNA insertion element and the polymerase chain reaction to amplify any M paratuberculosis DNA from intestinal tissue DNA extracts. One oligonucleotide primer was fluorochrome-labelled and the presence of fluorescent amplified product was determined using an automated DNA sequencer with a computerised gel-scanning laser. This method was shown capable of detecting 1-2 mycobacterial genomes. Intestinal tissue samples were obtained from 68 patients with histologically confirmed Crohn's disease, 49 patients with histologically confirmed ulcerative colitis, and 26 non-inflammatory bowel disease controls. In no case was M paratuberculosis detected in any of the inflammatory bowel disease tissue samples and only one non-inflammatory bowel disease case was positive. These results do not support the hypothesis that M paratuberculosis has an aetiological role in Crohn's disease.     

NP30/抗体检测试剂盒在云南大山区型血吸虫病流行区的现场应用

目的用NP30/抗体检测试剂盒在大山区型血吸虫病流行区筛查病人,评价其应用价值。方法在云南省大山区型血吸虫病流行区,选择3个自然村作为现场,随机抽取10～70岁的村民作为调查对象,进行血吸虫病病史个案调查;收集粪便,作毛蚴孵化检查;静脉采血,分离血清,分别采用NP30/抗体检测试剂盒和酶联免疫吸附试验试剂盒（SEA/ELISA法）检测特异性IgE和总IgE抗体。结果3个村共筛查村民506人,粪便毛蚴孵化阳性164例,阳性率为32.41%;NP30和SEA/ELISA的阳性率分别为58.30%和75.10%。在164例粪检阳性血清样本中,NP30和SEA/ELISA的阳性符合率分别为87.80%和84.76%。在342例粪孵阴性血清样本中,NP30和SEA/ELISA的阳性率分别为44.15%和70.47%。结论NP30/抗体检测试剂盒具有较高的敏感性和特异性,可用于大山区型血吸虫病流行区病人筛查。     

Comparison of stool microscopy and serology (enzyme linked immunosorbent assay) in epidemiology of amebiasis

Stools from 634 individuals from Varanasi were examined for Entamoeba histolytica (EH). Serology was done in these subjects by enzyme linked immunosorbent assay (ELISA) employing filter paper technique. Stools were positive for EH in 16.9%, and serology in 15.9%. Both the tests were positive in only 5.2%. In 72.4% both the tests were negative. In 11.7% of stool positive cases, serology was negative, and in 10.7% with positive serology stool examination did not reveal EH. A majority (92.5%) of stool positive subjects had only cysts. Additional parasites were detected in 15.3%.     

T-cellular immune reactions (in macrophage inhibition factor assay) against Mycobacterium paratuberculosis, Mycobacterium kansasii, Mycobacterium tuberculosis, Mycobacterium avium in patients with chronic inflammatory bowel disease.

A mycobacterial aetiology has been suggested for Crohn's disease. A slow growing mycobacterium, biochemically and genetically identical to M paratuberculosis, the causative agent of enteritis in ruminants (Johne's disease), has been isolated from gut specimens of patients affected by Crohn's disease. If M paratuberculosis or other mycobacteria play a role in the pathogenesis of Crohn's disease, then patients may have been sensitised to these mycobacteria or show an anergy immune reaction. We therefore investigated the T-cell mediated immune response to sonicates of M paratuberculosis, M kansasii, M avium, and M tuberculosis in 35 patients with Crohn's disease, 28 with ulcerative colitis, and 25 controls using a macrophage inhibition factor assay on peripheral blood lymphocytes. Two types of reaction patterns were identified--that is, 'responders' (subjects with a macrophage inhibition factor assay in which a dose response relation was present and a percentage of inhibition exceeding 20%), and 'non-responders'. There was no significant difference in the prevalence of responders (59%-80%) and non-responders (20%-41%) to these mycobacteria between the group of Crohn's disease, ulcerative colitis, and control group. We found also that a large proportion of controls showed T-cell immunisation to the mycobacteria which supports the contention that the antigens are practically commensal. Our results do not support the proposed involvement of mycobacteria in the pathogenesis of Crohn's disease.     

The obligate anaerobic faecal flora of patients with Crohn's disease and their first-degree relatives

The obligate anaerobic faecal floras of patients with Crohn's disease, their first-degree relatives, and healthy control subjects were compared. The flora of Crohn's patients contained more anaerobic gram-positive coccoid rods and gram-negative rods than that of healthy subjects; on this basis patients and healthy subjects formed two clusters with minor overlap. Nine of 26 children of Crohn's patients were also included within the Crohn's disease cluster. During 5 to 7 years of follow-up study 3 of them presented with remitting abdominal pain, diarrhoea, or weight loss, and in 1 of them Crohn's disease was diagnosed; none of the 17 children with a normal flora showed symptoms possibly due to Crohn's disease. It is concluded that the abnormal flora may be indigenous to subjects predisposed to Crohn's disease, suggesting a direct or indirect relationship between the abnormal faecal flora and Crohn's disease.     

Role of faecal calprotectin as non-invasive marker of intestinal inflammation

BACKGROUND/AIM: Faecal calprotectin, a neutrophil granulocyte cytosol protein, is considered a promising marker of intestinal inflammation. We assessed and compared the faecal calprotectin concentration in patients with organic and functional chronic intestinal disorders. PATIENTS AND METHODS: The study was carried out, using a commercially available ELISA test, measuring calprotectin in stool samples collected from 131 patients with inflammatory bowel diseases, 26 with intestinal neoplasms, 48 with irritable bowel syndrome and 34 healthy subjects. RESULTS: Median faecal calprotectin was significantly increased in Crohn's disease (231 microg/g, 95% confidence interval (CI) 110-353 microg/g), ulcerative colitis (167 microg/g, 95% CI 59-276 microg/g), and neoplasms (105 microg/g, 95% CI 0-272 microg/g), whereas normal values were found in patients with irritable bowel syndrome (22 microg/g, 95% CI 9-35 microg/g) and in healthy subjects (11 microg/g, 95% CI 3-18 microg/g). A positive correlation was observed with clinical activity scores in Crohn's disease and ulcerative colitis. In both groups, patients with clinically active disease showed higher calprotectin levels than those observed in patients with quiescent disease (405 microg/g, 95% CI 200-610 microg/g vs. 213 microg/g, 95% CI 85-341 microg/g in CD patients, p<0.05, and 327 microg/g, 95% CI 104-550 microg/g vs. 123 microg/g, 95% CI 40-206 microg/g in UC patients, p<0.001). CONCLUSIONS: Faecal calprotectin appears to be a promising and non-invasive biomarker of intestinal inflammation. If these findings are confirmed, it may provide a useful test for the diagnosis and follow up of inflammatory bowel diseases.     

Immunologic tests in patients after clinical cure of visceral leishmaniasis

The results of five serologic tests (ELISA using promastigote antigen [ELISAp] and recombinant K39 [ELISArK39] and K26 [ELISArK26] antigens, indirect immunofluorescence test using promastigote antigen [IIFT], and immunochromatographic tests using the rK39 antigen [TRALd]) and of the Montenegro skin test (MST) were analyzed in 41 individuals treated for kala-azar and living in Porteirinha, Minas Gerais, Brazil. The tests were carried out 1 week to 12 years after specific treatment. All MSTs during the 8 months after treatment were negative, whereas after 1 year, 28 (84.8%) were positive. Negativity in all serologic tests was observed for 11 (26.8%) of the 41 individuals, whereas positivity in at least one test was observed for 70.3% of subjects evaluated > or = 2 years after treatment. With respect to each exam, positivity was 38.0% for TRALd, 61.9% for ELISA rK39, 47.6% for ELISA rK26, 38.0% for ELISAp, and 40.5% for IIFT. None of the individuals presented recurrence of the disease during the 4 years of follow-up. The tests were repeated in 24 of the 41 individuals, after some time, and the results were the same in 33.3% of the cases. We conclude that serological tests for kala-azar might continue to be positive after treatment of the disease, although this does not indicate a poor prognosis or a poor therapeutic response.     

Evaluation of surgical tissue from patients with Crohn's disease for the presence of Mycobacterium avium subspecies paratuberculosis DNA by in situ hybridization and nested polymerase chain reaction

Crohn's disease (CD) is a chronic inflammatory bowel disease (IBD) with tissue granuloma and histopathological alteration that resembles aspects in tuberculosis, leprosy, and paratuberculosis. Mycobacterium avium subsp paratuberculosis (MAP) is the causative agent of paratuberculosis, with a suspected role in the etiology of CD. We investigated the presence of MAP DNA in 31 surgical tissue samples from 20 subjects using fluorescence in situ hybridization (FISH) with the aid of confocal scanning laser microscopy and nested polymerase chain reaction (PCR) using the IS900 sequence unique to MAP. MAP DNA was detected by PCR in tissue from 10 of 12 (83%) patients with CD: 7/12 (58%) in inflamed, 6/11 (55%) in noninflamed and in 10 (83%) of either tissue and by FISH in 8 of 12 (67%) patients with CD: 7 of 12 (58%) in inflamed, 4 of 11 (36%) in noninflamed, and in 8(67%) of either tissue. In non-IBD subjects, MAP DNA was detected in the tissue of only 1 of 6 patients (17%) by PCR and 0 of 6 patients (0%) by FISH. MAP DNA was identified by PCR in inflamed tissue from 2 of 2 patients with ulcerative colitis. The detection of MAP DNA by either technique in tissue from subjects with CD is significant compared with non-IBD subjects (P < 0.005). Identification of MAP DNA in both inflamed and noninflamed tissue by both techniques suggests that MAP infection in patients with CD may be systemic. The data add more evidence toward a possible association of MAP in the pathogenesis of CD.     

Isolation of peptides useful for differential diagnosis of Crohn's disease and ulcerative colitis

BACKGROUND: Phage displayed random peptide technology has been utilised to identify binding epitopes of antibodies or receptor ligands. Aim: To isolates peptides from a phage library which are specifically recognised by antibodies in serum from patients with Crohn's disease (CD). METHODS: A phage displayed random peptide library composed of nine amino acids was established and sequentially screened using serum immunogloblin G obtained from CD patients. RESULTS: Five different CD specific peptides were isolated from the phage library. No homology in amino acid sequences was observed among four (CDP-1, -3 to -5) of the five peptides exhibiting different binding characteristics with each CD patient's serum. In contrast, two peptides (CDP-1 and -2) had similar amino acid sequences and similar binding characteristics. Four multiple antigenic peptides (MAP, CDP-1, -3 to -5) were synthesised, and an enzyme linked immunosorbent assay (ELISA) using the four peptides was developed to detect serum antibodies against them. Fifty two of 92 CD patients (56.5%) were detected by ELISA, none of 20 ulcerative colitis (UC) patients, only one of 25 duodenal ulcer patients, and only three of 48 healthy subjects. CONCLUSIONS: ELISA using the four peptides isolated in this study may be useful for the differential diagnosis of CD and UC.     

Structural heterogeneity of faecal alpha 1 antitrypsin shown by immunoblot analysis in patients with Crohn's disease.

Faecal alpha 1 antitrypsin was determined in 34 patients with Crohn's disease and in 19 healthy subjects by immune nephelometry. A structural analysis of faecal alpha 1 antitrypsin was carried out using immunoblot analysis under non-reducing conditions. Native serum alpha 1 antitrypsin migrated with an apparent molecular weight of 45 kDa. Proteolytic alpha 1 antitrypsin fragments (5-42 kDa) were specifically immunostained in 13/19 and 22/34 stool samples from control subjects and from patients with Crohn's disease respectively. There was a weak correlation (r = 0.47; p less than 0.02) between the molecular weight of fragmented alpha 1 antitrypsin and the faecal concentration in both groups, indicating that alpha 1 antitrypsin inhibits its own proteolysis by intestinal proteases in a dose dependent way. The incidence of polymeric forms (greater than 45 kDa) was similar in patients (10/34) and control subjects (5/19). In only one case in each group was the native serum form of alpha 1 antitrypsin found in faeces. We conclude that faecal alpha 1 antitrypsin differs structurally from the native serum form. Immunochemical measurements, therefore, reflect rather than represent faecal concentrations of alpha 1 antitrypsin. The controversial results in published reports may be partly explained by these findings. The molecular heterogeneity of faecal alpha 1 antitrypsin is not specifically associated with Crohn's disease.     

Serum antineutrophil cytoplasmic autoantibodies in inflammatory bowel disease are mainly associated with ulcerative colitis. A correlation study between perinuclear antineutrophil cytoplasmic autoantibodies and clinical parameters, medical, and surgical treatment.

Perinuclear antineutrophil cytoplasmic antibodies have recently been demonstrated in the sera of patients with inflammatory bowel disease. Three hundred and sixty six sera obtained from 120 patients with ulcerative colitis, 105 patients suffering from Crohn's disease and 49 non-inflammatory bowel disease controls were tested in two laboratories, using an indirect immunofluorescence assay. In addition, a fixed-neutrophil enzyme linked immunoadsorbent assay (ELISA) was evaluated in one of the two laboratories. The results in the immunofluorescence test showed a high degree of correlation between the two laboratories (Kappa coefficient = 0.8). Ninety five of the 120 (79%) ulcerative colitis patients had a positive test whereas only 14 of the 105 (13%) patients with Crohn's disease were positive. Sera from four patients suffering from primary sclerosing cholangitis were positive as well as four of the 45 control sera (9%). The sensitivity of the perinuclear antineutrophil cytoplasmic antibody immunofluorescence test for the diagnosis of ulcerative colitis was 0.75 with a specificity of 0.88 and a positive predictive value of 0.88 (all sera). In the ELISA technique 37 of 94 ulcerative colitis sera and one of the 68 Crohn's disease sera were positive. In the control group only one of the patients suffering from primary sclerosing cholangitis reacted positively (32 non-inflammatory bowel disease sera tested). The ELISA technique had a high specificity (0.97), but a low sensitivity (0.39). There was no relation of perinuclear antineutrophil cytoplasmic antibodies in ulcerative colitis patients or in Crohn's disease patients with disease activity, duration of illness, localisation, extent of disease, previous bowel operations or medical treatment. The clinical significance of perinuclear antineutrophil cytoplasmic antibody positive and negative subsets in both groups of patients thus remains unexplained. Our study confirms that determination of serum antineutrophil cytoplasmatic antibodies in patients with inflammatory bowel disease may differentiate ulcerative colitis from Crohn's disease. Further immunological studies are needed to explain the absence of these antibodies in a subset of ulcerative colitis patients and their role in the pathogenesis of the disease.