Development of a quantitative-competitive PCR for quantification of human cytomegalovirus load and comparison with antigenaemia, viraemia and pp67 RNA detection by nucleic acid sequence-based amplification.

AIM: The human cytomegalovirus (HCMV) is an important pathogen in immunocompromised patients, such as transplant recipients. The use of sensitive and rapid diagnostic assays can have a great impact on antiviral prophylaxis and therapy monitoring and diagnosing active disease. Quantification of HCMV DNA may additionally have prognostic value and guide routine management. The aim of this study was to develop a reliable internally-controlled quantitative-competitive PCR (QC-PCR) for the detection and quantification of HCMV DNA viral load in peripheral blood and compare it with other methods: the HCMV pp65 antigenaemia assay in leukocyte fraction, the HCMV viraemia, both routinely employed in our laboratory, and the nucleic acid sequence-based amplification (NASBA) for detection of HCMV pp67-mRNA. METHODS: Quantitative-competitive PCR is a procedure for nucleic acid quantification based on co-amplification of competitive templates, the target DNA and a competitor functioning as internal standard. In particular, a standard curve is generated by amplifying 10(2) to 10(5) copies of target pCMV-435 plasmid with 10(4) copies of competitor pCMV-C plasmid. Clinical samples derived from 40 kidney transplant patients were tested by spiking 10(4) copies of pCMV-C into the PCR mix as internal control, and comparing results with the standard curve. RESULTS: Of the 40 patients studied, 39 (97.5%) were positive for HCMV DNA by QC-PCR. While the correlation between the number of pp65-positive cells and the number of HCMV DNA genome copies/mL and the former and the pp67mRNA-positivity were statistically significant, there was no significant correlation between HCMV DNA viral load assayed by QC-PCR and HCMV viraemia. CONCLUSIONS: The QC-PCR assay could detect from 10(2) to over 10(7) copies of HCMV DNA with a range of linearity between 10(2) and 10(5) genomes.

[The environment and health protection of the child]. / Ambiente e tutela della salute del bambino.

The authors outline the relationship between environment and children health care. They describe the environmental situations able to produce damages to the children's health.

[Goldenhar's syndrome: a case report]. / La sindrome di Goldenhar: descrizione di un caso osservato.

The Authors describe one case of Goldenhar's syndrome. They concentrate on the etiopathogenetic hypothesis and they believe it helps the gradual healing in time.

[Margagni-Larrey diaphragmatic hernia. Description of a clinical case]. / Ernia diaframmatica di Morgagni-Larrey. Descrizione di un caso clinico.

A clinical case report of a Morgagni-Larrey hernia is presented. The finding was made at the age of four months by means of x-ray of the thorax, performed because of recurrent respiratory symptoms regressed after surgical intervention. The rarity of this type of hernia, without a herniary sac and accompanied by the late appearance of respiratory symptomatology, is stressed. It is suggested that x-ray of the thorax should be performed when recurrent respiratory disease is the presenting complaint, even in the first few months of life.

[Gastric ulcer in the infant: clinical contribution]. / L'ulcera gastrica del lattante: contributo clinico.

The authors, after a short review on the etiopathological classification and the clinical exordium of the gastric ulcer in infancy, report a relapsing case nevertheless the therapy with cimetidine and antacid drugs. Furthermore they note the importance of the careful anamnesis and early and repeated occult blood stool tests to value for eventual gastric fiberendoscopy.

[Symptomatic Kimmerle anomaly. Clinical case]. / Anomalia di Kimmerle sintomatica. Caso clinico.

After mentioning the most common forms of vertigo in childhood, the Authors quote a case come into their observation showing anomalies of the Kimmerle symptomatic.

[The hematuria symptom. Behavior of the pediatrician]. / Il sintoma "ematuria." Comportamento del pediatra.

The authors have reviewed the symptom hematuria in childhood, suggesting a simple flow chart for ambulatorial pediatricians.

[The Guillain-Barré syndrome in childhood]. / La sindrome di Guillain-Barré nell'infanzia.

The authors describe a case day Guillain-Barré syndrome admitted in pediatrics hospital department for asthenia. Approximately an hour after admission, the patient showed dysphagia and respiratory insufficiency and was diagnosed a Guillain-Barré syndrome, successfully treated.

[A case of hereditary angioedema]. / Su di un caso di angioedema ereditario.

The authors describe a case of hereditary angioedema in a female child of four years. Treatment with C1 human-immuno-inactivator. Good success of therapy.

[Rheumatic manifestations and dental foci: a review of the cases in 12 years of activity in a pediatrics division]. / Manifestazioni reumatiche e foci dentari: revisione della casistica di 12 anni di attività in una divisione di pediatria.

By examining 8244 clinical records, in a period of 12 years of paediatric activity, the authors point out the connection between dental caries and rheumatic fever. They suggest fluoride supplementation since the early age, in countries where the fluoride is lack in drink able-water.

[Axial polydactyly: a clinical case]. / Polidattilia assiale: caso clinico.

After a short review of the normal embryonal development of lower limbs the authors describe a case of polydactyly associated with a 2nd-grade hypospadias observed in a 4-month-age patient. The authors, after the surgical treatment of both defects, have made the follow-up of the patient until six year age.

[A case of persistent fever]. / Su di un caso di febbre persistente.

The Authors report a case of difficult diagnosis, clinically characterised by long-term persistent fever. Clinical and laboratory tests led to a first conclusion of a tubercular infection; later of a perisplenic hematoma and the persistence of high inflammatory indices directed to a different diagnosis.

[Abdominal pain in childhood: multidisciplinary approach. A clinical case]. / Il dolore addominale in età pediatrica: approccio multidisciplinare. Caso clinico.

The Authors describe a case of mature cystic teratoma found in a little girl 9 + 8/12 years old and removed in laparoscopic way. Various specialists' contribution shows up for the resolution of the case.

[Cheilognathopalatoschisis: its embryology, epidemiology, etiology, clinical picture and therapy. The experience of a division of pediatrics]. / La cheilognatopalatoschisi: embriologia, epidemiologia, etiologia, clinica e terapia. Esperienza presso una divisione di pediatria.

The authors report their experience about cleft lip and palate.