Evaluation of the immunochromatographic (Leptocheck) test for detection of specific antibodies against leptospires.

BACKGROUND: Leptospirosis is a febrile worldwide zoonosis. Routine diagnosis of leptospiral infection is based on demonstration of specific antibodies with serological tests. Performance of the reference serological test, the microscopic agglutination test (MAT), requires significant expertise. The aim of our study was to find out if leptospiral infection can be proven with simple, rapid, commercially available immunochromatographic Leptocheck test in order to introduce it for the first level diagnosis in emergency cases with less specialized laboratory staff. METHODS: In all, 590 serum samples of patients with clinical manifestations suggestive of leptospirosis were collected and tested with MAT and Leptocheck test. For confirmation of the results some other diagnostic methods such as polymerase chain reaction (PCR) and Leptospira isolation were performed. RESULTS: Results of both serological tests were consistent in 576/590 (97.63%) cases but Leptocheck gave more positive results in comparison to MAT (36 and 12, respectively) at first patient's testing. Following up the patient, MAT became positive in majority of Leptocheck positive patients at first visit. Leptospiral DNA was detected in nine blood and six urine samples belonging to thirteen different patients while only two samples were culture positive. CONCLUSION: In comparison with serological tests, PCR and culture have low sensitivity. According to our findings we conclude that Leptocheck test can prove leptospiral infection and could be used for rapid diagnosis of leptospirosis, later the sample should be confirmed with MAT.

Impact of added fluvastatin to standard-of-care treatment on sustained virological response in naïve chronic hepatitis C Patients infected with genotypes 1 and 3.

OBJECTIVES: The combination of pegylated interferon-α and ribavirin is a standard-of-care (SOC) treatment for chronic hepatitis C (CHC), and it achieves a sustained virological response (SVR) in 41-52% of genotype 1 and in 73-79% of genotype 3 patients. In a few clinical trials, the combination of fluvastatin and SOC increased the SVR in genotype 1 patients. METHODS: This prospective study enrolled 179 naïve CHC patients. In the fluvastatin group patients received the combination of SOC and fluvastatin 80 mg daily; historical controls matching the study group in genotype, age and gender were treated with the SOC treatment only. RESULTS: On-treatment viral responses as well as the SVR did not differ significantly between the two groups, except for the genotype 1 patients with a high viral load presenting a significantly higher SVR rate in the fluvastatin group (75%) compared to the control group (41%; p = 0.024). Multivariate logistic regression identified hepatitis C virus (HCV) genotype 3 infection (p < 0.001), age ≤40 years (p < 0.001), liver steatosis <5% (p < 0.01) and low viral load (p < 0.001) as independent predictors of an SVR. CONCLUSION: A combination of fluvastatin and SOC significantly improved the SVR in naïve CHC patients infected with HCV genotype 1 and high viral load, but it did not improve the SVR in patients infected with HCV genotype 3.

Hepatitis C virus genotypes in 1,504 patients in Slovenia, 1993-2007.

In order to identify the main routes of hepatitis C (HCV) transmission and to determine the HCV genotype distribution and its dynamics during a 15-year period in Slovenia, HCV genotypes were detected using the INNO-LiPA HCV II (Innogenetics) test for serum samples obtained from 1,504 patients representing 72.6% of all patients with chronic hepatitis C diagnosed from 1993 to 2007. HCV genotype 1 was predominant (56%), followed by genotypes 3, 2, and 4, with a prevalence of 37.8%, 5%, and 1.2%, respectively. HCV genotypes 5 and 6 were not detected in any patient. Patients infected with HCV genotype 3 were significantly younger (mean age 28.9 +/- 8.5 years) than those infected with genotype 1 (mean age 38.9 +/- 14.8 years; P < 0.0001) and those infected with HCV genotype 2 (mean age 50.3 +/- 18.2 years; P < 0.0001). Intravenous drug use was identified as the most frequent possible HCV transmission route (34.3%), followed by medical-related transmission such as transfusion of HCV-contaminated blood or blood products, and hemodialysis (12.5%). Being an intravenous drug user was found to be strongly associated with HCV genotype 3 (OR, 3.71 [95% CI, 2.97-4.65]; P < 0.0001) and reporting infection by transfusion of blood or blood products was found to be strongly associated with HCV genotype 1 (OR, 3.28 [95% CI, 2.18-4.95]; P < 0.0001). During the 15-year period, the proportion of genotype 3 increased substantially, reflecting the fact that the HCV epidemic in Slovenia is driven mostly by intravenous drug use.

Temporomandibular joint involvement caused by Borrelia Burgdorferi.

BACKGROUND: Lyme borreliosis is an endemic disease in Slovenia with an incidence of around 150 patients per 100,000 inhabitants. Although the large joints are most typically affected in Lyme borreliosis, there are also periods of disease activity with arthritis or arthralgias involving smaller joints, including the temporo-mandibular joint. PATIENTS: During the years between 2000 and 2003, two patients with Lyme borreliosis affecting the temporo-mandibular joints were treated. The patients presented with fatigue and pain in diverse muscle groups accompanied by arthralgia, which was most pronounced in the temporomandibular joint area. None of the patients were febrile or had joint effusions. METHODS: Both patients were examined by means of biochemical and serological examinations for Borrelia burgdorferi using ELISA assay and Western blot test (both for IgM and IgG), plain radiographs, MR and CT scans, and scinti-scan of the temporo-mandibular joints They both had positive serum markers for an acute B. burgdorferi infection and were treated with intravenous ceftriaxone. RESULTS: None of the patients had clinical or laboratory signs of chronic Lyme disease activity two and four years following therapy, respectively. Roentgenographic and nuclear magnetic resonance imaging of the temporo-mandibular joints had not shown any persistent sign of acute inflammation. CONCLUSION: There are only few reports of patients with manifest temporo-mandibular joint involvement of Lyme borreliosis in the literature. This report emphasizes the importance of differential diagnosis of acute temporo-mandibular joint arthralgia, of early diagnosis of Lyme borreliosis, and of the necessity for prompt antibiotic treatment.

Accidental poisoning with autumn crocus.

We describe a case of a 43-yr-old female with severe multiorgan injury after accidental poisoning with Colchicum autumnale, which was mistaken for wild garlic (Allium ursinum). Both plants grow on damp meadows and can be confused in the spring when both plants have leaves but no blossoms. The autumn crocus contains colchicine, which inhibits cellular division. Treatment consisted of supportive care, antibiotic therapy, and granulocyte-directed growth factor. The patient was discharged from the hospital after three weeks. Three years after recovery from the acute poisoning, the patient continued to complain of muscle weakness and intermittent episodes of hair loss.

Pediatric tick-borne encephalitis in 371 cases from an endemic region in Slovenia, 1959 to 2000.

BACKGROUND: With the exception of Lyme borreliosis, tick-borne encephalitis (TBE) is the most prevalent tick-transmitted disease in Europe. Here we report clinical and epidemiologic features of the largest number of children with TBE reported to date and the longest (i.e. 42-year) retrospective survey of pediatric TBE cases from one geographic region. METHODS: Case records of 371 patients, age 0 to 15 years, with serologically confirmed TBE and hospitalized between 1959 and 2000 at the Department of Infectious Diseases of the General Hospital Celje, Slovenia were reviewed and analyzed. RESULTS: Children represented 23.5% of 1578 confirmed TBE cases in the study period. Children were admitted to hospital throughout the year, but the majority were treated during summer months. In 178 (47.9%) children, a tick bite was noticed before admission. A biphasic course of illness occurred in 249 (67.1%) patients. The most common symptoms and signs of TBE were raised body temperature [>38 degrees C (n = 371)], headache and meningeal signs (n = 346), fatigue (n = 337) and vomiting (n = 327). Meningitis was diagnosed in 232 (62.5%) children, and meningoencephalitis was diagnosed in 139 (37.5%). There was a tendency for greater severity of TBE with increasing age. None of the children with TBE died, and none had permanent sequelae. CONCLUSIONS: The results of our study indicate that pediatric TBE is relatively mild disease with favorable outcome.

Does end-stage kidney failure influence hepatitis C progression in hemodialysis patients?

BACKGROUND/AIMS: The influence of end-stage kidney failure on the progression of liver disease in patients infected with hepatitis C virus and treated with hemodialysis is still controversial. METHODOLOGY: Liver histology of 154 hepatitis C virus infected non-uremic patients was compared with liver histology of 13 hepatitis C virus infected uremic patients treated with hemodialysis. RESULTS: In either group of altogether 167 patients, no normal liver histology was found. Each patient had at least a low-grade lobular and/or portal inflammation. However, statistically significant differences were observed between hepatitis C virus infected uremic and non-uremic patients in the extent of intralobular changes, portal inflammation, and degree of fibrosis. CONCLUSIONS: Non-uremic hepatitis C virus infected patients appear to have more active and progressive liver disease than hepatitis C virus infected patients on hemodialysis. Regular follow-up of uremic patients, associated with earlier detection of hepatitis C virus infection, so as suggested uremia-associated impaired immunoreactivity and increased levels of hepatocyte growth factor described recently, might be implicated in a more favorable course of hepatitis C virus infection in uremic patients. In addition, due to the absence of normal liver histology in either group of hepatitis C virus infected patients, we propose liver biopsy to be mandatory in all these patients, provided that no contraindications exist clinically.

Efficacy of chronic hepatitis C therapy with interferon alpha (IFN-alpha) in Slovenia.

BACKGROUND/AIMS: Eighty Slovene patients with chronic hepatitis C were included in a prospective study conducted in the period 1997-1998 with the purpose to establish the efficacy of interferon alpha therapy. The average age of the patients was 39 years. In more than half of the patients (52%) the mode and time of onset of the infection were unknown. Two thirds of the patients were males. The plasma viral load exceeded 2 x 10(6) copies/mL in only three patients and in more than half of the cases (54%) HCV genotype 1b was present. METHODOLOGY: The 18-month treatment with 3 MU interferon alpha three times a week was concluded in 53 patients and, after doubling the initial dose of interferon alpha from 3 MU to 6 MU, in 5 patients. In 11 patients, the treatment was discontinued prematurely, after six months, due to therapeutic failure (despite doubling the initial dose of interferon alpha Eleven patients withdrew from the treatment: six of them due to side effects and five due to personal reasons. RESULTS: Complete response to therapy with disappearance of HCV from the blood was observed in 34 patients (49%), while in 24 the response to therapy was partial, i.e., the biochemical tests showed normalization of values but viremia persisted. There was a significant relation between the therapeutic response and those patients with the genotype 3 (p = 0.01). After three months of follow-up, complete therapeutic response was still observed in 19 patients (28%), most of them with genotype 3. Despite persistent viremia there was no progression of liver inflammation in eight partial responders, as evidenced by liver rebiopsy. Thus, it was confirmed that treatment is justified in these patients. CONCLUSIONS: During the continuation of the follow-up period we shall record further course of the disease and make an attempt in a subsequent study to improve the efficacy of the treatment by introducing a combination of interferon alpha and ribavirin into therapy.

Comparative evaluation of three commercial assays for quantitative measurement of hepatitis B virus DNA in serum samples.

BACKGROUND/AIMS: Quantitative determination of HBV DNA in serum samples is indispensable for predicting disease progression and for monitoring the antiviral treatment in patients with chronic HBV infection. METHODOLOGY: Three commercial assays for quantification of HBV DNA: Digene Hybrid-Capture HBV DNA Assay, Bayer Quantiplex HBV DNA Assay and Roche Amplicor HBV Monitor Test were comparatively evaluated under the routine conditions of diagnostic virology laboratory, using 61 serum samples obtained from 55 Slovenian patients with chronic hepatitis B. RESULTS: HBV DNA was detected by Amplicor, Quantiplex and Hybrid-Capture in 38 (62.3%), 34 (55.7%) and 27 (44.3%) samples, respectively. The sensitivity of Amplicor and Quantiplex assays did not differ significantly (p = 0.13), while both Amplicor and Quantiplex assays were found to be significantly more sensitive than Hybrid-Capture (p = 0.003 and p = 0.02, respectively). For a given sample, the highest correlation was observed between HBV DNA loads determined by Quantiplex and Hybrid-Capture assays (r = 0.85, p < 0.0001). CONCLUSIONS: Amplicor HBV Monitor Test seems to be the most sensitive assay for the detection of HBV DNA in serum samples and can be clinically used for monitoring patients with chronic HBV infection.