Real-time PCR for detection of low intensity Schistosoma japonicum infections in a pig model.

Decades of successful Schistosoma japonicum control have increased the interest in how to diagnose low intensity infections. A real-time PCR assay targeting the mitochondrial NADH dehydrogenase I gene in S. japonicum was evaluated in infected pigs with very low egg output. Six out of 12 S. japonicum infected pigs were treated with praziquantel 8 weeks after infection and all pigs were followed for 16 weeks post-infection. One commercial and one non-commercial extraction method were evaluated in combination with PCR on faecal samples. PCR with either extraction method were equally sensitive as the DBL-filtration/sedimentation technique in the acute, productive stage. PCR recovered slightly more positive samples in the chronic stage, but most faecal samples were negative for both PCR and microscopy from week 9 post-infection irrespective of treatment. IgG antibody titers against soluble egg antigen IgG remained high throughout the study in both the treated and non-treated group. PCR was consistently negative in serum and urine samples and negative in most of the caecal biopsies. We conclude that the S. japonicum faecal PCR is a highly sensitive test. However, in clinical samples when faecal egg output almost reaches nil in the chronic stage despite persistent worm burdens, both the faecal PCR and microscopy results were negative. Real-time PCR is less labour intensive than most microscopy methods, but has a higher material cost per sample.

Novel real-time PCr for detection of Schistosoma japonicum in stool.

Chemotherapy has been used on a large scale in countries where the blood fluke Schistosoma japonicum is endemic. This has led to a lower intensity of infections and consequently lower diagnostic values of commonly used diagnostic tests like serology and Kato-Katz stool smear. We designed a novel real-time PCR method for detection of S. japonicum in stool samples. Further, we evaluated different versions of an inexpensive, non-commercial extraction method, ROSE, as well as the commercial QIAamp DNA Stool Mini Kit. PCR primer sequences were designed targeting the mitochondrial NADH dehydrogenase I gene. Bovine serum albumin was added to the DNA extracts and SYBR Green was used for detection. The PCR method was evaluated with non-infected stool samples spiked with S. japonicum eggs. It demonstrated high sensitivity, even in samples containing a single egg. The two extraction methods were equally effective. The PCR was specific for S. japonicum when tested against other Schistosoma species, Trichuris trichiura, hookworm and Taenia sp. We conclude that this novel real-time PCR, in combination with either ROSE or QIAamp DNA Stool Mini Kit extraction, is a sensitive and specific tool for diagnosing S. japonicum in human stool samples.

Antibiotic sensitivity still prevails in Norwegian blood culture isolates.

We describe the antimicrobial susceptibility of bacteraemia isolates from Norway. From March 1998 to February 1999, four university hospitals covering all parts of Norway collected their first 10 isolates each month. Minimal inhibitory concentrations were determined for: Enterobacteriaceae (n=192), staphylococci (n=89) and Streptococcus pneumoniae (n=69) using the Etest. NCCLS breakpoints were used. About 20% of all blood culture isolates in Norway in this period were investigated. Compared with countries outside Scandinavia antibiotic sensitivity still prevails. Only minor differences in resistance were found between participating hospitals, between hospital departments and between hospital- and community-acquired pathogens. The prudent use of antibiotics in Norway may contribute to the fact that antibiotic resistance still remains low in the most common bacterial pathogens causing bloodstream infections.

Complete right bundle branch block after surgical closure of perimembranous ventricular septal defect. Relation to type of ventriculotomy.

The ECGs of a 100 consecutive children who had surgical repair of their ventricular septal defects (VSDs) were analyzed for postoperative right bundle branch block (RBBB). Seventy of them had an atriotomy and the other 23 also a ventriculotomy. The ventriculotomy always consisted of a transverse incision a short distance below the pulmonary annulus. Of these children 93 had a perimembranous VSD and the other 7 a pure muscular defect. The ECG results of the 93 children with perimembranous VSDs were statistically analysed. The incidence of postoperative complete RBBB (CRBBB) in the ventriculotomy group was not higher than in the atriotimy group. Infants operated in the first half year of life were more prone to the development of CRBBB than the older children, probably because the VSDs were relatively larger in the younger than the older children. The risk of postoperative CRBBB was less in the children who had direct suture closure of the VSD compared with those who needed a Dacron patch to close the defect. The data in the literature generally indicate a higher incidence of CRBBB after a ventriculotomy than an atriotomy. The absence of this difference and the lower incidence of CRBBB after a ventriculotomy in our series compared with those of several other authors are suggested to be due to the type of ventriculotomy.

Pharmacokinetics and body distribution of amiodarone in man.

The concentrations of amiodarone (Cordarone) and desethylamiodarone in plasma after single oral and intravenous and long-term oral dosing were determined in seven normal subjects and 106 patients with various cardiac arrhythmias, respectively, using a high-performance liquid chromatographic method. The decline in amiodarone plasma concentration after a single intravenous 400 mg dose was described by a triexponential decay equation, with a mean terminal half-life (t1/2) of 34.5 h. Model independent parameters were calculated from the fits. Mean values for clearance and apparent volume of distribution were 14.7 +/- 7.2 l/h and 376 +/- 372 l. Following single oral doses of 400 mg, amiodarone plasma concentration time data were fitted in a triexponential function. The mean terminal half-life for amiodarone after oral dosage was 31.6 +/- 21.3 h. Amiodarone peak concentrations of 0.37 +/- 0.22 micrograms/ml were attained in 4.8 +/- 1.5 h. The bioavailability of oral amiodarone was only 31 +/- 26%, in part due to first-pass metabolism. Desethylamiodarone , the major metabolite of amiodarone, was present in plasma in very low levels of about 10 ng/ml in several volunteers after single intravenous or oral administration of amiodarone. The mean plasma amiodarone and desethylamiodarone levels in 106 patients, using a mean oral daily maintenance dosage of 440 +/- 253 mg for a mean period of 9.1 months, were 1.85 +/- 1.17 micrograms/ml and 1.35 +/- 0.71 micrograms/ml, respectively. The relationship between the steady state amiodarone and desethylamiodarone plasma concentrations and daily amiodarone maintenance dose in mg in 106 patients was investigated.(ABSTRACT TRUNCATED AT 250 WORDS)