Emergency extracorporeal life support and ongoing resuscitation: a retrospective comparison for refractory out-of-hospital cardiac arrest.

BACKGROUND: In refractory cardiac arrest, with cardiopulmonary resuscitation (CPR) for more than 30 min, chances of survival are small. Extracorporeal cardiopulmonary resuscitation (ECPR) is an option for certain patients with cardiac arrest. The aim of this study was to evaluate characteristics of patients selected for ECPR. METHODS: Anonymised data of adult patients suffering refractory cardiac arrest, transported with ongoing CPR to an ED of a tertiary care centre between 2002 and 2012 were analysed. Outcome measure was the selection for ECPR. Secondary outcome was 180 days survival in good neurological condition. RESULTS: Overall, 239 patients fulfilled the inclusion criteria. ECPR was initiated in seven patients. Patients treated with ECPR were younger (46 vs 60 years; p=0.04), had shorter intervals before CPR was started (0 vs 1 min; p=0.013), faster admissions at the ED (38 vs 56 min; p=0.31) and lower blood glucose levels on admission (14 vs 21 mmol/L; p=0.018). Survival to discharge in good neurological condition was achieved in 14 (6%) of all patients. One patient in the ECPR group survived in excellent neurological condition. Age was independently associated with the selection for ECPR (OR 0.07; 95% CI 0.01 to 0.85; p=0.037). CONCLUSIONS: Emergency extracorporeal life support was used for a highly selected group of patients in refractory cardiac arrest. Several parameters were associated with the decision, but only age was independently associated with the selection for ECPR. The patient selection resulting in a survival of one patient out of seven treated seems reasonable. Randomised controlled trials evaluating the age limit as selection criteria are urgently needed to confirm these findings.

[Structures and Practice of Psychological Services in Oncological and Diabetological Rehabilitation: Results of a Nationwide Survey]. / Strukturen und Praxis der psychologischen Abteilungen in der onkologischen und diabetologischen Rehabilitation: Ergebnisse einer bundesweiten Befragung.

OBJECTIVE: To study the structural frame conditions and the contents of psychological activity in oncological rehabilitation as well as in rehabilitation of patients with type 2 diabetes. METHODS: We conducted a nationwide survey of psychological services in rehabilitation facilities treating oncological patients and patients with type 2 diabetes. RESULTS: 71 (of 145) oncological and 21 (of 63) diabetological rehabilitation facilities participated in the survey. In both indication areas an average of 1.1 psychologists is in charge of 100 patients. Between some rehabilitation facilities, however, there are considerable differences concerning the psychologist/patient ratio (in oncological rehabilitation facilities: standard deviation (SD)=0.52; in diabetological rehabilitation facilities: SD=0.35). Moreover, there is large heterogeneity among rehabilitation facilities as to the percentages of patients obtaining psychological interventions and the way in which psychological services allocate their working time. CONCLUSION: The general set-up of psychological services in oncological and diabetological rehabilitation facilities (especially the low psychologist/patient ratio in many facilities) can partly be considered insufficient. The heterogeneity with respect to the structural frame conditions and practice of psychological services reveals the low degree of standardization of psychological activity in both indication areas.

Morphometric evaluation of the linea alba in fresh corpses.

Purpose: Diastasis recti abdominis is an increase in the distance between the medial borders of the two rectus muscles. It is most often triggered after intra-abdominal pressure increases, such as postpartum or in obesity. Most publications are based on radiological studies or are done in certain subgroups, without unanimous reference values of the distance between the rectus abdominis or standardization. Methods: Forty-one cadavers were studied. Exclusion criteria: signs of abdominal trauma, major burns, presence of scar from previous abdominal surgery, clinical signs of abdominal hernia, and identification of hernia during cadaver dissection. Linea alba (LA) length, width, and thickness were measured with a flexible tape measure and digital caliper. Anatomical landmarks were established, and subdivisions were described based on them to compare the cadavers. Results: Sex and age had little effect on LA width, thickness, or length. Obesity (compared to normal weight) was the only variable that promoted an increase in the LA width (p < 0.01). The supraumbilical length varied with the total height of the evaluated cadavers (p < 0.01), but the infraumbilical length did not (p = 0.11). Conclusion: The general statistical results of this study, regarding the evaluation of LA measurements in cadavers, showed that ethnicity, sex, and age have little effect on the width, thickness, or length of the LA. LA width differed significantly with abdominal circumference.

[DGRW-Update: Rehabilitation in Diabetes Mellitus]. / DGRW-Update: Rehabilitation bei Diabetes mellitus.

In the years to come, prevalence and socio-medical relevance of diabetes mellitus will continue to increase. Therapeutic aims must be defined on an individual basis considering risks and benefits. No longer is it reasonable to insist on normoglycaemia as a general therapeutic aim. There are numerous effective and evidence-based therapeutic modules for diabetes mellitus which are also offered within the scope of rehabilitation. Reliable evidence exists to confirm that therapy should start as early as possible, because it is less effective during later phases of the disease when concomitant cardiovascular illnesses may occur. In most cases, medical rehabilitation of diabetic patients is based on other diagnoses. There is a considerable need for rehabilitation among diabetics who are in ambulant care but do not intend to file a request for rehabilitative measures. Sustainability of rehabilitative effects must be improved by means of follow-up treatment and networking with the ambulant structures of long-term care. Provided that the indication makes it appropriate, bariatric surgery constitutes a new effective therapy.

[Fractionated inpatient rehabilitation of diabetes: results from a randomized controlled trial on rehabilitation aftercare]. / Intervallrehabilitation bei Diabetes mellitus: ergebnisse einer randomisierten kontrollierten Studie zur Nachsorge in der medizinischen Rehabilitation.

BACKGROUND: Treating diabetes mellitus type 2 (DMT2) essentially involves long-term changes of health behaviour, especially diet and physical activity habits. Medical rehabilitation tries to support these changes by patient education and practical instructions. While short-term successes are frequently obtained, the main challenge lies in maintaining these results in the longer run. This study examines whether a fractionated inpatient rehabilitation programme of 3 weeks duration in combination with an additional week of inpatient rehabilitation after 6 months and a subsequent aftercare telephone service facilitates positive long-term effects for patients with DMT2. METHOD: A prospective randomized controlled trial was conducted with 411 patients with DMT2 in order to compare the fractionated inpatient rehabilitation programme with aftercare to a 3-weeks standard rehabilitation programme without aftercare. The analysis included physiological (cardiovascular risk, HbA1c, BMI) and psychosocial (diabetes-specific coping, quality of life) parameters at baseline (beginning of the programme) and 1 year later. RESULTS: After 12 months, no significant differences between the groups were found in the physiological parameters. At the same time, however, the aftercare group showed larger benefits in almost every psychosocial dimension (coping, quality of life, subjective health). Coping and quality of life had even improved when compared to baseline. In contrast, these parameters had decreased further in the standard-care control group when compared to baseline. CONCLUSION: The intensive aftercare was successful in the long-term improvement of the patients' psychological well-being. Also, the high rate of participation in fractionated inpatient rehabilitation suggests both the acceptance of and the need for aftercare. However, positive long-term changes of physiological parameters probably will require more specific interventions or individual case management. The effectiveness of fractionated inpatient rehabilitation may be limited by the adverse social, psychological and financial situation of the patients.

Imbalance between plasma double-stranded DNA and deoxyribonuclease activity predicts mortality after out-of-hospital cardiac arrest.

AIM: Despite an increased rate of return of spontaneous circulation (ROSC) in out-of-hospital cardiac arrest (OHCA) patients, almost half of patients do not survive up to hospital discharge. Understanding pathophysiological mechanisms of post-cardiac arrest syndrome is essential for developing novel therapeutic strategies. During systemic inflammatory responses and concomitant cell death, double-stranded (ds) DNA is released into circulation, exerting pro-inflammatory effects. Deoxyribonuclease (DNase) degrades dsDNA. The role of DNase activity in OHCA survivors and impact on clinical outcome has not been analyzed yet. METHODS: In a prospective, single-center study, dsDNA and DNase activity were determined at hospital admission (acute phase) and 24 h (subacute phase) after ROSC. The ratio between dsDNA levels and DNase activity was calculated to determine the extent of dsDNA release in relation to the patients' capacity of degradation. Thirty-day mortality was defined as study end point. RESULTS: We enrolled 64 OHCA survivors, of whom 26.6% (n = 17) died within 30 days. A peak of circulating dsDNA was observed at admission which decreased within 24 h. DNase activity did not differ between acute and subacute phase, while dsDNA load per DNase activity significantly decreased. The ratio between dsDNA levels and DNase activity in the subacute phase was the strongest predictor of 30-day mortality with an adjusted HR per 1 SD of 3.59 (95% CI, 1.80-7.18, p < 0.001). CONCLUSION: Disproportionally increased dsDNA levels uncompensated by DNase activity are a strong predictor of mortality in OHCA survivors. This pilot study points to a potentially protective effect of DNase activity in patients undergoing cardiac arrest.

The Subcutaneous Heparin and Angioplasty Restenosis Prevention (SHARP) trial. Results of a multicenter randomized trial investigating the effects of high dose unfractionated heparin on angiographic restenosis and clinical outcome.

OBJECTIVES: We sought to determine whether 12,500 IU of unfractionated heparin given subcutaneously twice daily for 4 months after percutaneous transluminal coronary angioplasty beneficially influences the subsequent rate of angiographic restenosis and the incidence of clinical events. BACKGROUND: Heparin has been shown to exhibit powerful antiproliferative effects against smooth muscle cells in several animal models. METHODS: A randomized trial with blinded data analysis was undertaken to assess the effect of unfractionated subcutaneous heparin on angiographic restenosis after coronary angioplasty. After successful angioplasty, patients were randomized to receive no heparin or 12,500 IU of heparin given subcutaneously twice daily for 4 months. Quantitative coronary angiography was performed before angioplasty, immediately after angioplasty and at follow-up ("early" [before 4 months] or electively [at 4 months]). RESULTS: The study group comprised 339 patients, 167 randomly assigned to receive heparin, 172 to receive no heparin. Repeat cardiac catheterization was performed in 90% of randomized patients. At early and elective restudy (mean 4.2 months), the mean +/- SD difference in minimal lumen diameter between the postangioplasty and follow-up measurement was -0.55 +/- 0.58 mm for the no heparin group and -0.43 +/- 0.59 mm for the heparin group (p = NS). Clinical events during the follow-up period did not differ significantly between groups: fatal myocardial infarction (1 patient in each group), coronary bypass grafting (5 patients in each group), repeat angioplasty (12 in the no heparin, 6 in the heparin group), angina at 4-month assessment (33% in the no heparin, 32% in the heparin group). CONCLUSIONS: Long-term treatment with high dose subcutaneous heparin (12,500 IU twice daily) for 4 months did not favorably influence angiographic or clinical outcome after coronary angioplasty.

Bent DNA is needed for recombinational enhancer activity in the site-specific recombination system Cin of bacteriophage P1. The role of FIS protein.

A series of recombinational enhancer mutants was constructed by manipulating the ClaI site between the two FIS binding sites of the Hin enhancer. These mutants include insertions from two to 12 base-pairs and two deletions of one or two base-pairs. Recombinational enhancer activity was found only with four mutants carrying either a four base-pair substitution, ten base-pair insertions or a one base-pair deletion, respectively; two other ten base-pair insertion mutants, however, were inactive, although FIS protein binding was unaffected. So, besides binding of FIS protein to its specific sites within the enhancer sequence and the correct helical positioning of these sites on the DNA, another criterion for enhancer activity must be fulfilled. DNA bending assays identify this requirement as a change of the enhancer DNA conformation, which FIS protein is able to induce and to stabilize. This conformational change of the DNA can be blocked by mutations in the central segment between the two FIS binding sites of the Hin enhancer. This sequence has special functions for the recombinational enhancer activity.

Random mutagenesis using degenerate oligodeoxyribonucleotides.

An efficient method for random mutagenesis was applied to a 75-bp target sequence. The mutational changes in the target region are introduced by the technique of oligodeoxyribonucleotide(oligo)-directed, site-specific mutagenesis using mixtures of degenerate oligos. These are designed in such a way that they carry with a high probability randomly distributed substitutions, which are introduced into the oligos by utilizing appropriate concentrations of all four nucleotide precursors during each chain elongation step. These mixtures of degenerate oligos were hybridized to the appropriate M13-hybrid ss-template and then extended in vitro using PolIk. In order to avoid any bias artificially created by the Escherichia coli mismatch repair system, homoduplex molecules were synthesized in vitro according to the method of Taylor et al. [Nucleic Acids Res. 13 (1985) 8765-8785]. After transformation of the appropriate E. coli host, M13 plaques were randomly analysed by DNA sequencing. Using appropriate preparations of template DNA and oligos we attained mutagenesis efficiencies in the range of 20-50%. The analysis of 85 different mutants revealed that the distribution of the mutations is random and that all expected substitutions occur with about the same probability.

Sequence and transcript analysis of the nitrogenase structural gene operon (nifHDK) of Rhodobacter capsulatus: evidence for intramolecular processing of nifHDK mRNA.

Northern blot analysis of RNA prepared from cells of Rhodobacter capsulatus derepressed for nitrogenase (N2ase) synthesis, using a 6.0-kb DNA probe containing the entire nifHDK operon, revealed the presence of at least six hybridizing species of the estimated sizes, 4.4, 3.5, 2.7, 1.3, 0.9 and 0.38 kb. No hybridization was detected with RNA prepared from cells grown in the presence of an excess of NH4+, which represses N2ase synthesis. Hybridization with gene-specific probes revealed that the 4.4-kb species hybridized with all three probes, and presumably corresponded to the full-length nifHDK transcript, whereas the 3.5-kb species hybridized with nifD and nifK only, and the 2.7-kb transcript hybridized with nifH and nifD. The 1.3 and 0.9-kb species hybridized with all three probes, but appeared to hybridize most strongly with nifH. In contrast, the 0.38-kb species hybridized with none of the gene-specific probes, and was also detected in RNA from cells of strain RcM1, which contains a chromosomal deletion of the nifHDK operon. This species probably corresponds to the transcript of a gene, named fdxD, which was found to be located just upstream from the nifHDK operon. Nucleotide (nt) sequencing of the nifH-D and nifD-K intergenic regions revealed the presence of inverted repeat (IR) sequences potentially capable of forming stable stem-loop structures in mRNA. Primer extension analysis of the nifDK-homologous species showed that the 5' end was located one or two nt downstream from the IR sequence between nifH and nifD, suggesting that the putative stem-loop structure may be a target for intramolecular processing of the nifHDK mRNA.

Study of the presence of Campylobacter jejuni and C. coli in sand samples from four Swiss chicken farms.

Chicken farms are frequently infected with Campylobacter jejuni and Campylobacter coli. The objective of the present study was to investigate environmental samples from chicken farms for the presence of C. jejuni and C. coli. Every week between July and November 1997, three sand samples from the runs of four chicken farms were analyzed by culture and directly by polymerase chain reaction (PCR). These two detection methods were compared to each other. A total of 231 samples were tested. Eleven samples (4%) were found to contain Campylobacter cells by culture, whereas 157 samples (68%) were positive by PCR. All samples which were positive by culture were also positive by PCR. All direct PCR products were further typed by restriction fragment length polymorphism (RFLP). Three different RFLP types and mixtures of these types were observed. Direct PCR products of one chicken farm were further typed by direct sequencing and two temporally separated sequence types could be distinguished. Campylobacter strains isolated by culture were also typed by RFLP and direct sequencing revealing close accordance with the corresponding direct PCR products.

PCR-based detection of verotoxin-producing Escherichia coli (VTEC) in ground beef.

Pathogenic strains of Escherichia coli producing verotoxins (VTs) have been recognized as a cause of human disease, and rapid and sensitive detection tests are urgently needed to ensure the safety of food, especially ground beef. We applied two nested polymerase chain reaction (PCR) assays to detect the genes encoding VT1 and VT2 irrespective of the bacterial serotype. In combination with a direct sample preparation protocol, we were able to uncover the presence of about 110 CFU of verotoxinogenic E. coli (VTEC) in 10 g of ground beef. When a six-hour enrichment was included, we found the detection limit to be in the range of 1 to 10 bacterial cells per 10 g of ground beef. To evaluate our detection system, we tested 30 ground beef samples originating from butcher shops in Berne, Switzerland. One sample yielded positive PCR results for both the VT1 and VT2 genes, indicating the presence of verotoxinogenic E. coli. Finally, 20 food homogenates, shown to contain E. coli strains by standard culture, were analysed with our method, and the gene encoding VT2 was detected in one cheese sample. The results suggest that the described PCR method can serve as a valuable tool for the surveillance of VTEC contamination of foods.

Analysis of benzalkonium chloride in the effluent from European hospitals by solid-phase extraction and high-performance liquid chromatography with post-column ion-pairing and fluorescence detection.

A highly reproducible and specific method for the analysis of the quaternary ammonium compound, benzalkonium chloride, in effluents from European hospitals is presented. Benzalkonium chloride was extracted with end-capped RP-18 solid-phase cartridges and was selectively eluted. The resulting solution was analyzed by high-performance liquid chromatography (HPLC). After elution from the analytical column of the HPLC system, 9,10-dimethoxyanthracene-2-sulfonate was added continuously as a fluorescence marker, forming a hydrophobic ion-pair with benzalkonium chloride. The ion-pair was analyzed by fluorescence detection. The method was applied to highly complex effluent samples from different sized European hospitals. The measured concentrations were between 0.05 and 6.03 mg/l. The amounts emitted per bed and year were 4.5-362 g and did not correlate with the size of the hospital. The total amounts were 2.6-909 kg/year.

Assessment of myocardial perfusion and function using gated methoxy-isobutyl-isonitrile scintigraphy to detect restenosis after coronary angioplasty.

BACKGROUND: Technetium-99m-methoxy-isobutyl-isonitrile (MIBI) has been shown to be a useful perfusion agent, and its improved imaging characteristics also allow evaluation of myocardial function. We evaluated the use of gated MIBI scintigraphy in patients undergoing percutaneous transluminal coronary angioplasty to show improvements in myocardial perfusion and function and to detect restenosis. METHODS: Gated MIBI scintigraphy was performed in a preliminary study in 25 patients before angioplasty. Twenty-one patients who had undergone a successful procedure were re-examined 3 months later. Myocardial perfusion was evaluated and function measured using myocardial profiles along four axes. RESULTS: Before angioplasty, 18 out of 21 patients had a reversible perfusion defect, and computer analysis of fractional shortening showed 19 out of 21 had abnormalities of left ventricular function during exercise. Only 11 out of 21 had a positive stress test. Mean global fractional shortening before angioplasty was 27.3% at rest and 26.6% during exercise (not significant). After angioplasty, left ventricular function was improved during exercise, with global fractional shortening increasing from 26.6% to 32.7% (P < 0.001). Repeat angiography was performed in 16 patients. Lesions in four patients had restenosed, in 11 they had not, and in one a different artery was occluded. Three of the four patients with restenosed lesions were identified using repeat perfusion imaging, and all four from abnormalities in left ventricular function. An abnormality in a new region was detected using both methods in the patient with the new occlusion. Normal perfusion and function were found in nine of the 11 patients without restenosed lesions. Two patients continued to have a positive stress test: one with a restenosed lesion and one with new disease. CONCLUSION: Dual assessment with gated MIBI scintigraphy provides useful data in patients undergoing angioplasty and may help in the selection of patients for repeat angiography.

Gated technetium-99m methoxy-isobutylisonitrile perfusion imaging.

Technetium-99m methoxy-isobutylisonitrile, has been shown to be a useful perfusion agent for detecting coronary artery disease. Gated acquisition of perfusion images may remove motion artefact and improve detection. We compared the results of sensitivity and specificity for detecting coronary artery disease using perfusion images from the whole cardiac cycle (ungated) or from end-diastolic (gated) frames in 46 subjects, 31 with significant coronary disease and 15 with normal coronary arteries. There was no significant difference in detection of patients with coronary disease between the ungated and end-diastolic images: 25/31 with the ungated image and 28/31 with the end-diastolic. An improvement was made in detecting defects in the left anterior descending artery territory: 17/20 with the end-diastolic vs 11/20 with the ungated image (P less than 0.05) and in the right coronary artery region: 16/21 vs 10/21 (P less than 0.05), with an overall improvement in detecting regions supplied by stenosed coronary arteries: 45/55 end-diastolic vs 31/55 ungated (P less than 0.01). These results suggest use of the end-diastolic frame can significantly enhance the diagnostic capacity of methoxy-isobutylisonitrile perfusion imaging.

Monitoring anticoagulation following intracoronary procedures: which method?

We have assessed bedside kits for monitoring the activated partial thromboplastin time and the activated clotting time by comparing them with laboratory activated partial thromboplastin time values. To determine the accuracy of anticoagulation we have concurrently measured the plasma heparin concentrations, and plasma prothrombin fragment F1 + 2 concentrations. Serial samples were taken from patients undergoing elective percutaneous transluminal coronary angioplasty (n = 14). Readings were taken pre-procedure, 30 min after administration of a heparin bolus (10,000 U) and 1, 2 and 3 h after commencement of a constant heparin infusion (15 U/kg/h) postprocedure. Activated partial thromboplastin time results obtained with the bedside kit compared reliably with laboratory values (r = 0.8), were rapidly available and were reflected by appropriate changes in prothrombin fragment F1 + 2 and heparin concentrations. However, the relationship between activated partial thromboplastin time values and activated clotting time was less precise (r = 0.59). Therefore, for routine and frequent monitoring of anticoagulation with heparin, a bedside activated partial thromboplastin time kit provides adequate control of therapy but in instances were particularly tight control of anticoagulation is required, use of prothrombin fragment F1 + 2 concentrations may be more appropriate.

Prothrombin fragment F1 + 2 concentrations for monitoring anticoagulation therapy with heparin.

We have compared the activated partial thromboplastin time with measurement of prothrombin fragment F1 + 2 concentrations (ELISA assay) during a 24-h period in a group of patients (n = 10) who had undergone elective coronary angioplasty and were anticoagulated post-procedure with heparin 1000 U/h. Four hours after the procedure all the patients were adequately anticoagulated according to activated partial thromboplastin time (median ratio 4.7:1) and the prothrombin fragment F1 + 2 concentration was significantly lower than pre-angioplasty values (0.5 vs 1.4 nmol/l, p = 0.04). At 24 h the median activated partial thromboplastin time ratio was still higher than the pre-procedure value (1.35 vs 0.9, p < 0.01), but the prothrombin fragment F1 + 2 concentration had risen to 2.1 nmol/l, with more variability in individual results within the patient group for the prothrombin fragment F1 + 2 concentration than for activated partial thromboplastin time (interquartile ranges (Q1, Q3) prothrombin fragment F1 + 2, 1.2-2.5; activated partial thromboplastin time, 1.2-1.5). The activated partial thromboplastin time is the standard method of monitoring the anticoagulant effect of heparin but may not fully reflect the functional coagulation status and accurately identify individual patients with less than adequate anticoagulation. Prothrombin fragment F1 + 2 concentrations may provide a more reliable indicator in individual patients of functional coagulation status in certain important situations where anticoagulation is critical such as following complicated coronary angioplasty.

Long term use of enalapril in the treatment of patients with congestive heart failure.

Fifty-five patients of both sexes, aged 39-72, suffering from congestive heart failure of varying aetiology were admitted to a double blind study in which enalapril given in a dose of 5-10 mg twice daily or placebo was added to existing medication. A significant increase in exercise performance (P less than 0.004) was seen in the group treated with enalapril, an improvement which was still evident after 24 weeks of therapy. No significant difference was seen between the mean ejection fractions but subjective assessment suggested improvement in the enalapril-treated group.

Outbreak of viral gastroenteritis due to sewage-contaminated drinking water.

In August 1998, a large outbreak of gastroenteritis occurred in a Swiss village of 3500 inhabitants whereof more than 50% were affected. A high contamination of drinking water with faecal coliforms revealed a defect in the waste water system. The objective of the present study was to investigate the outbreak in respect of the presence of human pathogenic viruses. Drinking water and clinical samples from patients were examined for the presence of 'Norwalk-like viruses' (NLVs) and enteroviruses. NLVs and enteroviruses were detected by reverse transcription-polymerase chain reaction (RT-PCR) in one of two drinking water samples. Five of seven stool samples from ill persons were positive for NLVs. Typing of NLV-specific RT-PCR products by DNA sequencing revealed the presence of an identical genogroup-1 strain closely related to Southampton virus in both the water and one of the stool samples. A genogroup-2 NLV strain was identified in all positive stool samples. The enteroviral amplicon showed high sequence similarity with swine vesicular disease virus. These results demonstrate that the drinking water was highly contaminated with enteric viruses and that at least two NLV strains were involved in this outbreak of gastroenteritis.

Three-step isolation method for sensitive detection of enterovirus, rotavirus, hepatitis A virus, and small round structured viruses in water samples.

Control of drinking or bathing water quality in respect to viral contamination remains an unsolved problem. A highly sensitive isolation protocol was developed for concentration and detection of different enteric viruses from water samples. The three-step isolation procedure combines filtration with a positively charged nylon membrane, ultrafiltration and clean-up of the viral RNA with a silica based membrane. Detection of the viral RNA is accomplished by reverse-transcription polymerase chain reaction (RT-PCR). Detection limits were determined to be one 50% tissue culture infective dose (TCID50) of seeded coxsackievirus B2 or hepatitis A Virus per litre of tap water by RT-PCR compared to two orders of magnitude lower sensitivity for culture in the case of coxsackievirus B2. The isolation procedure is highly sensitive, easy to perform and allows the detection of different human pathogenic virus groups in one water sample. The application of the isolation procedure to six river water samples and subsequent detection with nested or semi-nested PCR revealed enterovirus in 6/6 (100%), rotavirus in 6/6 (100%), hepatitis A virus in 0/6 (0%), small round structured virus genotype I in 6/6 (100%) and small round structured virus genotype II in 2/6 (33%) of the samples. These findings suggest that first, we have developed a very sensitive detection procedure and second, that river water in Switzerland-where most of the wastewater is handled by sewage treatment plants-shows a high contamination rate with enteric viruses.