CMV gB genotypes and outcome of vertical transmission: study on dried blood spots of congenitally infected babies.

BACKGROUND: The role of the virulence of the infecting cytomegalovirus (CMV) strain in the transmission of the virus from mother to fetus and the outcome of the fetal infection has not received much attention yet. Molecular analysis of the gene coding for the surface glycoprotein B (gB) has been used to investigate the relationship between genotype and virulence in groups of immunosuppressed patients. OBJECTIVES: (1) to assess the prevalence of different gB genotypes in babies with congenital CMV infection; (2) to investigate the possible relationship between genotype and severity of congenital CMV disease; (3) to evaluate the possibility of using dried blood on Guthrie cards (DBS) for genotyping. STUDY DESIGN: CMV DNA was extracted from DBS and from urine/saliva samples collected in the first two weeks of life of 98 congenitally infected babies, half of which were symptomatic at birth. Genotyping was performed through RFLP analysis of the region corresponding to the cleavage site of the gB protein. RESULTS: The most prevalent genotype was gB1 (42%) followed by gB3 (26%), gB2 (19%) and gB4 (13%). Rates of disease and CNS damages were higher among children infected by gB1 (35%, 17%) and gB3 (31%, 28%) than in those infected by gB2 and gB4 (20%, 17% and 13%, 15%, respectively). These differences however did not reach the statistical significance. The parallel typing of DBS and urine/saliva strains gave a full concordance of results. CONCLUSIONS: All four major CMV gB genotypes (gB1-4) can cause a congenital infection but none seems to be associated to the development and the severity of disease. The possibility of using the neonatal DBS for genotyping opens a way to the examination of large numbers of cases of congenital CMV infection.

Cytomegalovirus DNA detection in Guthrie cards: a powerful tool for diagnosing congenital infection.

BACKGROUND: A simple and reliable diagnosis of congenital cytomegalovirus infection is necessary both for clinical and epidemiological purposes. This could be accomplished through the demonstration of cytomegalovirus (CMV) DNA in blood spots (DBS) on Guthrie cards. OBJECTIVES: (1) To assess the sensitivity and specificity of the method (DBS test) in diagnosing congenital CMV infection compared with viral isolation and (2) to evaluate the applications of the test to the late diagnosis of congenital CMV. STUDY DESIGN: The method was tested on the cards of (1) 509 babies examined through viral isolation within their third week of life (72 positive cases) and (2) 191 children studied after 3 weeks of life (25 days to 5 years). Blood was eluted from Guthrie cards and heat extracted. The products of a nested polymerase chain reaction (PCR) amplifying one region in the CMV glycoprotein B (gB) gene were detected by agarose gel electrophoresis. RESULTS: DBS test was positive in all 72 congenitally infected babies and in four of the 437 negative at cytomegalovirus isolation (sensitivity 100%, specificity 99%). Infection in 16 of the 92 infants with a late viral isolation was demonstrated to be congenital by the test, which also detected congenital infection in 18 of 83 children in whom viral culture was not performed (13 with and five without symptoms). Fifty-six additional control cases tested negative. CONCLUSIONS: DBS test is a reliable assay for diagnosing congenital cytomegalovirus infection and could be used as an alternative to viral culture. It is able to reveal whether ascertained CMV infection is congenital or postnatal at an age when viral isolation is not able to do so. It can assess the role of risky procedures such as transfusion and it can ascertain the etiology of morbid conditions diagnosed late or of controversial origin.

Micronuclei in lymphocytes of young patients under antileukemic therapy.

The micronucleus test in peripheral blood lymphocytes was employed in the cytogenetic monitoring of children with acute lymphocytic leukemia (ALL), who had undergone chemotherapy and radiotherapy. Patients were treated with a variety of drugs, which included vincristine, methotrexate, daunomycin and prednisone; they also underwent cranial irradiation at the end of the first intensive phase of therapy. The first group under study consisted of 15 subjects on therapy, who showed a marked increase in micronucleated lymphocytes (mean: 19.96 +/- 12.96%) as a consequence of treatment compared with the control group (mean: 3.67 +/- 1.55%), while lower average values were obtained from 15 other subjects at the end of treatment (mean: 13.16 +/- 8.44%). A group of 6 patients was monitored during the entire period of therapy, namely at diagnosis, after 3 months of therapy, throughout maintenance therapy and at the end of it. The whole treatment lasted about 2 years. The results revealed a marked increase in basal micronucleus frequency, due to therapy: the micronucleated lymphocyte frequency remained significantly high throughout the treatment for almost all patients. These data clearly suggest the validity of the methodology in pointing out the role played by antileukemic agents in inducing somatic genetic damage.

[Relations between HDL cholesterol and total cholesterol in serum of normal subjects]. / Rapporto fra HDL-colesterolo e colesterolo totale nel siero dei soggetti normali.

High-density lipoprotein (H.D.L.) cholesterol has been measured in 96 normal subjects. Females had significantly higher H.D.L. cholesterol than males in the control group: 60.8 /+- 13 mg/dl - 55.7 /+- 9.8 mg/dl.

[Lipid variations in coronary disease]. / Variazioni del quadro lipidico nella malattia coronarica.

Levels of total cholesterol (TC), triglyceride (Tg), high-density-lipoprotein-cholesterol (HDL-C), apolipoproteins A and B (ApoA and ApoB), were assayed in the serum of 63 patients suffering from coronary atherosclerosis, who survived myocardial infarction (M.I.) compared to healthy subjects. TC was assayed by an enzymatic-colorimetric technique: after precipitation with PEG 6000 the same method was used to determine HDL-C. Tg were assessed using total enzymatic method and ApoA and ApoB by RID. On the basis of TC, Tg, HDL-C, ApoA and ApoB values we have identified 6 main patterns, presenting a very characteristic lipaemic model. No significant difference was found between survivors and controls as regards TC and Tg; on the contrary, there was a statistical difference between the two groups for HDL-C, ApoA and ApoB.

[Immunochemical determination of immunoglobulins and lysozyme in normal human saliva - principal assay technics]. / Determinazione immunochimica delle immunoglobuline e del lisozima nella saliva umana normale - principi tecnici di determinazione.

We have studied a method to determine the concentrations of low level of immunoglobulins and lysozyme in saliva. The concentration of IgG, IgA and IgM in human whole saliva were measured by fluorimetric method and the concentration of lysozyme by turbidimetric method. The results of our study in 18 normal subjects (15 females and 3 males), have showed the geometric mean concentration (microgram/ml) IgG 5,1; IgA 177; IgM 2,6 and lysozyme 117.