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Aim: To evaluate the Helicobacter pylori stool antigen (HpSA) test in the assessment of H. pylori infection and the effect of omeprazole treatment on its accuracy. Methods: Study 1: 140 dyspeptic patients were enrolled in the study and defined as H. pylori positive if histology and rapid urease test, or culture alone were positive. HpSA was performed on all patients and 13C-urea breath test (UBT) on 87. Study 2: 75 patients testing positive using both UBT and HpSA, were given omeprazole 20 mg for 2 weeks (Group A) or omeprazole 40 mg for 2 weeks (Group B), or OAC for 1 week (group C). A Helicobacter pylori stool antigen test was performed on all patients on days 3, 5, 7 and 14 during treatment, and also on days 7 and 14 post-treatment in groups A and B. UBT was performed in groups A and B on days 7 and 14 during treatment, and days 7 and 14 post-treatment. Results: 80/140 patients were H. pylori positive. The sensitivity and specificity of HpSA were 93.8 and 90%, similar to UBT (93.9 and 92.1%). Omeprazole significantly reduced both HpSA and UBT values, resulting in a decreased accuracy. Of 25 patients receiving 20 mg omeprazole, HpSA gave 5 and 6 false negatives after 7 and 14 days treatment respectively, while UBT gave 4 and 7 false negatives after 7 and 14 days treatment. Of 25 patients receiving 40 mg omeprazole, HpSA gave 7 and 9 false negatives after 7 and 14 days of treatment, while UBT gave 8 and 9 false negatives after 7 and 14 days of treatment. Two weeks after stopping omeprazole treatment, the HpSA and UBT were positive in all cases. Conclusions: The Helicobacter pylori stool antigen test is valuable in the assessment of H. pylori infection. Short-term omeprazole treatment decreases the accuracy of both HpSA and UBT in a similar manner.

Our aim was to evaluate the accuracy of HpSA test in the diagnosis of Helicobacter pylori infection after the end of eradication therapy. In all 106 H. pylori-positive patients (55 men and 51 women, mean age 51 years, range 19–82) were treated with a course of eradicating regimen. [13C]Urea breath test (UBT) and HpSA were performed four weeks after stopping the treatment. The diagnostic accuracy of HpSA was evaluated in comparison with the results of [13C]UBT. In 90 patients (85%) H. pylori was eradicated according to [13C]urea breath test. After eradication, sensitivity of HpSA was 87.5%, specificity 95.5%, positive predictive value 77.8%, negative predictive value 97.7%, and diagnostic accuracy 94.3%. HpSA is a valuable test in the posteradication assessment of H. pylori infection.

Objective. To investigate whether the systemic antibody response to Helicobacter pylori heat shock protein B can be considered, in addition to anti cytotoxin-associated protein (CagA) antibody determination, a further serological marker of increased risk of gastric cancer development. Methods. A total of 98 Giemsa positive Helicobacter pylori patients (28 with gastric cancer, 30 with duodenal ulcer and 40 with non-ulcer dyspepsia) were studied. Serum samples obtained from all patients were tested for IgG antibodies to CagA (116 kDa), VacA (89kDa) and heat skock protein B (54 kDa) antigens of Helicobacter pylori by the Western blot technique. Results. 2628 patients (92.9%) with gastric carcinoma, 2930 patients (96.7%) with duodenal ulcer and 3040 patients (75.0%) with non-ulcer dyspepsia were seropositive for CagA protein. The prevalence of serum IgG antibody to CagA in the cancer patients was not significantly higher than in duodenal ulcer and non-ulcer dyspepsia patients. The prevalence of antibodies to VacA was not significantly different between gastric carcinoma and non-ulcer dyspepsia patients. In contrast the prevalence of systemic antibodies to heat skock protein B was significantly higher in gastric cancer patients (78.6%) than in duodenal ulcer (36.7%, p=0.002) or non-ulcer dyspepsia patients (52.5%, p=0.029). Conclusions. The detection of antibodies to heat shock protein B is proposed as an additional test which, in association with the determination of serum antibodies to CagA, could help in determining the risk of developing severe gastroduodenal disease, and gastric cancer, in particular.

Noninvasive tests for Helicobacter pylori are increasingly used. Recently, an enzyme immunoassay for H. pylori detection in feces has been put on the market. Aim of this multicenter study was to evaluate the usefulness of this novel test as a predictor of H. pylori status in the pretreatment setting. Three hundred consecutive patients were enrolled. None of the patients had received any eradicating treatment in the last 12 months, and all underwent gastroscopy with biopsies of the antrum and body for histology (H) and rapid urease test (RUT). H. pylori status was defined positive (or negative) if both H and RUT were positive (or negative). When H and RUT gave conflicting results, the patients were classified as H. pylori-indeterminate. A stool specimen was collected for each patient and tested by using a novel enzyme immunoassay for H. pylori detection (HpSAT). Sensitivity, specificity, and diagnostic accuracy of the test were calculated, as was the cost of each assay. H. pylori status was positive in 159 patients, negative in 131, and indeterminate in 10. HpSAT gave evaluable results (positive or negative) in 293 patients, and doubtful results in 7 (2.3%). Sensitivity, specificity, and diagnostic accuracy of HpSAT were 96.8%, 89.7%, and 93.6% respectively. Considering the H. pylori-indeterminate patients as positive, the percentages were 95.8%, 98.7%, and 93.2% respectively. The cost for each assay was about US $27. These results suggest that HpSAT is a noninvasive, simple, reliable, fast, and cheap method for evaluating H. pylori status in the pretreatment setting.

Background. BKV infection usually occurs in early childhood through the respiratory tract.The virus persists in a latent form in the kidney and it could be reactivated under favorable condition.The most important clinical manifestations affect kidney transplanted in which the BK polyomavirus nephropathy (PVAN) can lead to kidney failure. Objectives. The aim of our study was to evaluate the clinical utility of quantification of BKV viruria and especially viremia detected by real-time PCR method to select the patients at risk of PVAN. Study Design. We carried out a quantitative (dosing) assay of BKV-DNA in 24 patients transplanted in Salerno’s hospital, or elsewhere, all treated with cyclosporine or tacrolimus and mycophenolate mofetil or Prednisolone. The enrollment was made on the basis of impaired renal function, in particular of the values of serum creatinine (> 25% of baseline level) and / or appearance of proteinuria. The nucleic acid extraction was performed by EXTRAgen kit (Nanogen); the extracts were submitted to quantitative evaluation by BKV Q-PCR Alert Kit indicare regione target in Real Time PCR (Nanogen) using the instrument ABI7300 (Applied Biosystems). Results. 16 patients were negative both for viremia and viruria,4 patients showed positive viruria but viremia 10,000 copies / ml. In the last group, biopsy, performed to diagnose PVAN was positive and immunosuppressive therapy was reformulate leading to the decline, but never to the negativity of viral load. Conclusions. The renal impairement combined with the quantification of BKV’s viral load in urine and especially in plasma, can also be effective predictors of PVAN.

Introduction.We analyzed the clinical case of a 51 years old man, kidney transplanted on December 2002. On April 2011, he had acute rectal bleeding, renal chronic rejection (creatinine 2.9 mg/dl), Hgb 8.7 g/dl, positive anti-CMV antibodies (IgG). A colonoscopy showed diverticulosis of the rectum associated with deepithelialisation. The patient was treated with maintenance immunosuppressive post-transplant therapy. On June 2011, the colonoscopy showed a stenosing lesion of the sigmoid colon, and blood sampling and intestinal biopsy were performed to search Cytomegalovirus (CMV) DNA by PCR. Methods. The presence of CMV-DNA was sought by automatic extractor QIACUBE, using QIAamp DNA BLOOD Mini Kit (Qiagen) for whole blood and QIAamp DNA Mini Kit (Qiagen) for biopsy.The extracted DNA was then amplified by Real Time PCR using Q-CMV RealTime Complete Kit (Nanogen), on instrument Applied Biosystems 7300. Results. At disease onset the viral load in whole blood was 208000 Geq/ml, and biopsy was positive. Antiviral therapy with Ganciclovir led to the negativity of the viral load and remission of symptoms. Conclusions. The clinical case described presented a reactivation of CMV infection in the intestine after more than 10 years from kidney transplantation, while the highest incidence of CMV reactivation usually occurs during the first year. In our opinion, the reactivation can be traced to long-term immunosuppressive therapy (maintenance posttransplant therapy) in combination with a state of inflammation of the intestinal mucosa. In fact, patients with IBD treated with steroid drugs, in particular the group of refractory to therapy and thus have a recovery of the inflammatory process, are exposed to reactivation of CMV with intestinal localization.

Background.The relationship between CMV and inflammatory bowel disease (IBD) is not yet fully understood; the viral reactivation could be related to the prolonged steroid treatment of patients with ulcerative colitis (UC). Study Design. Our study was conducted on 25 patients with reactivation of UC, all were screened for CMV research. CMV DNA was carried out by automatic extraction in whole blood and intestinal biopsies and amplified by RealTime PCR. Objectives.We assessed the prevalence of CMV infection and clinical outcome, particularly in CMV positive patients treated with antiviral therapy. Results. CMV DNA was detected in 11/25 patients (44%), 7 positive in both blood and biopsies, 2 positive biopsies, 2 positive in blood, but low charge. 14 CMV DNA negative patients: 8 had remission with conventional therapy, 6 treated with IFX (of which 2 underwent colectomy). 11 CMV DNA positive patients: 7 had remission with conventional therapy, 3 treated with IFX+Ganciclovir, 1 with Ganciclovir, obtaining negative viral load and clinical remission. Conclusion. The literature data available on the association CMV - IBD are partly conflicting. The decision on treatment with antiviral therapy of CMV positive patients is based on course of disease. On the other hand, some patients had remission without antiviral therapy. We considered viral load and resistance to conventional therapy to admit patients into antiviral treatment, getting acute phase remission. There are many unanswered questions regarding management of CMV reactivation in UC patients, which need long-term follow-up studies and larger population.

Background. HCMV is the most opportunistic viral agent in the transplantation of bone marrow and solid organ. Early therapy based on the detection of HCMV pp65 antigen in peripheral blood leukocytes, has led to a significant reduction in the incidence of related HCMV diseases.The pp65 antigenemia is difficult to standardize, while the quantification of HCMV DNA by Real-Time PCR is an alternative diagnostic approach with greater sensitivity and reproducibility, providing important information in the management of infected patients. Objectives. The aim of this study was the comparative analysis of quantitative HCMV DNAemia in leukocytes and in whole blood, with Real-time PCR, in a BMT patient with HCMV post-transplant reactivation, in order to analyze the relationship between levels of viral DNA at different time-points obtained in the two biological matrices. Study Design. The presence of HCMV DNA was detected in whole blood and peripheral blood leukocytes in a patient who underwent allogeneic marrow transplant for Ph1 + acute lymphoblastic leukemia in molecular remission. After 5-6 months, the patient has increased molecular Bcr-Abl (Philadelphia chromosome). It was activated immune reaction by means of tapering (lowering the dose of cyclosporine) that uses the GvL effect to turn negative the Philadelphia chromosome positivity (Bcr/Abl negativity). Later, the patient develops GvHD and cortisone is administered. The persistence of grafts treated with immunosuppressants periodically reactivates HCMV infection. The DNA extraction from whole-blood was performed by automatic extractor QIACUBE (Qiagen), while extraction from leucocytes was performed on a standard number of leukocytes (EXTRAcell- Nanogen).The extracted DNA was amplified by Real-Time Alert Q-PCR (Nanogen).The samples were analyzed weekly for about 5 months from 1 year after transplantation. Results. The patient at 1 year after transplantation, has three HCMV reactivation at 56th, 62th and 67th week. In all reactivations a good overlap values of viraemia in both whole blood leukocytes is shown. Conclusions. Our data confirm that the whole blood is a biological matrix with diagnostic similar performance not only to pp65 antigenaemia, as supported by extensive scientific literature, but also to the DNAemia in leukocytes that has been for a long time considered the elective matrix for evaluating the post-transplant HCMV infection, for pre-emptive therapy particularly in BMT patients.