Analysis of 31 CFTR mutations by polymerase chain reaction/oligonucleotide ligation assay in a pilot screening of 4476 newborns for cystic fibrosis.

OBJECTIVES: Molecular biological testing for genetic diseases has grown rapidly, but speed, accuracy, specificity, sensitivity, throughput, and cost become more important as large scale screening is considered. This is a pilot study of an assay for the simultaneous detection of up to 31 cystic fibrosis mutations in a multicentre population based screening of 4476 Italian newborns. METHODS: The assay is a polymerase chain reaction, followed by an oligonucleotide ligation assay (PCR/OLA) and finally a sequence coded separation. It allows the detection of up to 31 mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Guthrie spots were used as a source of template DNA. RESULTS: 144 carriers were detected during the analysis of 4476 samples, which translates into a carrier frequency of 1/31.1. Forty two carriers were detected from 1341 samples in Pavia (1/31.9), 53 from 1574 in Turin (1/29.7), and 49 from 1561 in San Giovanni Rotondo (1/31.8). Fifteen different mutations were detected, the most common being delta F508 (0.625). Other common mutations included G542X (16 of 144), which was particularly common in southern Italy (14 of 49), N1303K (8 of 144), and R117H (8 of 144), detected only in the northern centres. CONCLUSIONS: PCR/OLA is a robust, accurate, user friendly method for cystic fibrosis screening of newborns using blood spots in a semiautomated way at a low cost per mutation (0.8 Euro).

A pilot C282Y hemochromatosis screening in Italian newborns by TaqMan technology.

Hereditary hemochromatosis (HH) is a disorder of iron metabolism that leads to iron overload in middle age and can be caused by homozygosity for the C282Y mutation in the HFE gene. Preliminary studies have estimated the frequency of this mutation at 0.5-1% in Italy, but this has not been verified on a large sample. We analyzed 1,331 Italian newborns for the C282Y mutation in the HFE gene using dried blood spots (DBS) from the Neonatal Screening Center in Turin, Italy. The mutation was assessed using a semi-automatable 5'-nuclease assay (TaqMan technology). We detected 55 heterozygotes and no homozygotes in our sampling, resulting in an overall frequency of 2.1% +/- 0.6 for the C282Y allele. Differences in allele frequency were observed, and ranged from 2.7% +/- 1.3 in samples from Northern Italy, to 1.7% +/- 0.9 in samples from Central-Southern Italy. The low frequency of the at-risk genotype for iron overload suggests that genetic screening for HFE in Italy would not be cost effective. The present study, in addition to defining C282Y frequency, documents detection of the major HFE mutation on routine DBS samples from neonatal screening programs using a semi-automatable, rapid, reliable, and relatively inexpensive approach.

[Diagnosis of bacterial infections of the urinary tract]. / La diagnostica delle infezioni batteriche delle vie urinarie

The aetiopathogenesis of bacterial infections of the urinary ways and the techniques used to isolate and identify the bacteria present in the urine are considered. The following aspects are covered: 1) Introduction (Clinical microbiology); 2) Urine and bacterial multiplication; 3) Diagnostic techniques in urinary way infections; 4) Urine leucocyte count; 5) Urine bacteria count; 6) Guidelines for applying bacteriological research; 7) Recurrent infections of the urinary ways; 8) Diagnosis of the site of infection; 9) Bacterial indentification and incidence.

Piedmont region (Italy): setting quality standards and testing.

In this report we describe the two years (1991-1992) EQA programmes organized by the health committee of the Piedmont region (Italy). This programme was mandatory for all public and private laboratories. The parameters analysed were: PT and APTT (for coagulation); Hb, RBC, WBC, platelets and MCV (for haematology); glucose, urea, creatinine, total proteins, cholesterol, uric acid, triglycerides, AST, ALT, LDH, GGT, amylase, alkaline phosphatase, sodium, potassium, calcium, chloride, phosphorus and iron (for clinical chemistry). For haematology, the comparison between the first and the second year shows a significant decrease of imprecision, especially for platelets, WBC and MCV.

[Requisition, reply forms and data recording (author's transl)]. / Accettazione, refertazione ed archiviazione.

A system for the automatic handling of laboratory data is described with particular reference to the microbiology module. The system is based on "real time" operation which permits acquisition of data as soon as they are generated; computation as soon as the necessary elements became available, production and storage of results as soon as computed.

[Bacterial meningitis. Study from 1970 to 1988 at the Regina Margherita Children's Hospital in Turin]. / Le meningiti batteriche. Indagine dal 1970 al 1988 presso l'Ospedale Infantile Regina Margherita di Torino.

From 1970 to 1988, 330 cases of bacterial meningitis were diagnosed at the Children's Hospital Regina Margherita of Turin. Neisseria meningitidis was the most frequent cause of bacterial meningitis (45%); meningitis caused by Haemophilus influenzae increased from 4% in 1970-1983 to 19% in 1984-1988, and meningitis with pathogen unknown decreased from 28% in 1970-1983 to 7% in 1984-1988. The neonatal meningitis in 1984-1988 were caused by Streptococcus B in 90% of cases.

Reference values for immunoglobulin kappa and lambda light chains and the kappa/lambda ratio in children's serum.

We analyzed 708 serum samples from healthy children and adolescents by immunonephelometry to obtain reference values for the immunoglobulin kappa (kappa) and lambda (lambda) light chains and for their ratio at a time of life when immunoglobulin synthesis is maturing and continually being stimulated. The lambda chain concentration that is to be maintained throughout the child's life is reached very early, just after 1 year, whereas the concentration of the kappa chains, which increases gradually, reflects the concentration of the immunoglobulins as a whole. These reference values may be useful for studying kappa and lambda chains in illnesses involving the immune system in children.

[Effectiveness of cystolusis with rifamycin SV in patients with permanent catheters]. / Efficacia della cistolusi con rifamicina SV in pazienti con catetere a permanenza.

It has tested the effect of Rifamicina SV in the cystolusis employed on patients with permanent catheter: compared to the treatment with physiological solution this one, with Rifamicina SV, has showed, in most cases, the lower incidence of urine bacteria and their quick negative answer. This by an only local theatment. During the experiment has also been showed the possibility for the Rifamicina SV of selecting the Citrobacter as a resistant bacterial species.

An outbreak of febrile gastroenteritis associated with corn contaminated by Listeria monocytogenes.

BACKGROUND: On May 21, 1997, numerous cases of febrile gastrointestinal illness were reported among the students and staff of two primary schools in northern Italy, all of whom had eaten at cafeterias served by the same caterer. METHODS: We interviewed people who ate at the cafeterias about symptoms and foods consumed on May 20. There were no samples of foods left at the cafeterias, but we tested routine samples taken on May 20 by the caterer and environmental specimens at the catering plant. The hospitalized patients were tested for common enteropathogens and toxins. RESULTS: Of the 2189 persons interviewed (82 percent of those exposed), 1566 (72 percent) reported symptoms; of these, 292 (19 percent) were hospitalized. Among samples obtained from hospitalized patients, all but two of the stool specimens and all blood specimens were negative for common enteropathogens. Listeria monocytogenes was isolated from one blood specimen and from 123 of the 141 stool specimens. Consumption of a cold salad of corn and tuna was associated with the development of symptoms (relative risk, 6.19; 95 percent confidence interval, 4.81 to 7.98; P<0.001). L. monocytogenes was isolated from the caterer's sample of the salad and from environmental specimens collected from the catering plant. All listeria isolates were serotype 4b and were found to be identical on DNA analysis. Experimental contamination of sterile samples of the implicated foods showed that L. monocytogenes grew on corn when kept for at least 10 hours at 25 degrees C. CONCLUSIONS: Food-borne infection with L. monocytogenes can cause febrile illness with gastroenteritis in immunocompetent persons.

Hematological studies in children with Down syndrome.

Previous studies have reported erythrocyte macrocytosis in adults and children with Down syndrome (DS), the significance of which remains unclear. We compared hematological parameters of 50 DS children aged 2 to 15 years, divided into three age groups, with those of 68 aged-matched healthy children. Patients with DS had a significantly increased mean corpuscular volume (MCV) and hemoglobin in all groups when compared with the controls. Erythrocyte creatine content, hexokinase (Hk) activity, erythrocyte and serum folates, vitamin B12, haptoglobin, serum iron, and ferritin were tested. All of these parameters were not significantly different from those of the control group. We conclude that macrocytosis may not be an expression of reduced red cell survival but rather of an altered folate remethylation pathway, secondary to enhanced cystathionine beta-synthase (CBS) activity, the gene for which is present on chromosome 21.

Differential effects of oral conjugated estrogens and transdermal estradiol on insulin-like growth factor 1, growth hormone and sex hormone binding globulin serum levels.

In postmenopausal women oral ethinylestradiol causes a reduction in circulating insulin-like growth factor 1 (IGF-1) and an increase in serum growth hormone levels. There are no data on the effect of conjugated estrogens, the preparation most often used in estrogen replacement treatment (ERT), on these parameters. We evaluated serum IGF-1 and growth hormone levels, together with the levels of sex hormone binding globulin (SHBG), an indicator of estrogen hepatocellular action, before and after 6 months of ERT in two comparable groups of postmenopausal women. Sixteen women were treated with oral conjugated estrogens, 0.625 mg/day, and 14 with transdermal estradiol, 0.05 mg/day. In the women treated with oral conjugated estrogens, an increase in SHBG (p < 0.001), a decrease in IGF-1 (p < 0.001) and an increase in growth hormone (p < 0.05) serum levels were observed. No such effects were seen with the use of transdermal estradiol, devoid of hepatocellular effects. Undoubtedly, oral conjugated estrogens, 0.625 mg/day, through a hepatocellular effect, cause marked modifications in the IGF-1/growth hormone axis, which may have clinical relevance. For instance, the decreased IGF-1 level, together with the increased level of SHBG, might provide some explanation of the favorable epidemiological data on breast cancer risk in women receiving oral conjugated estrogens.