A Novel Multiplexed Enzyme-Linked Immunosorbent Assay for the Detection of IgG Seroreactivity to Cytomegalovirus (CMV) UL144.

Infection with human cytomegalovirus (CMV) is common and may have grave consequences in transplant recipients and congenitally infected children. Diagnosis of CMV infection is based on detection of specific antibodies and molecular assays. The incorporation of CMV serological assays into diagnostic algorithms requires careful evaluation and interpretation. Very few serological assays measure CMV infection by a specific strain. We developed an enzyme-linked immunosorbent assay (ELISA) using CMV-encoded UL144 as the antigen. UL144 encodes three major genotypes, A, B, and C, and recombinants. The ELISA was developed with the three UL144 proteins and optimized as a multiplex assay. Sera from 55 positive and 59 negative CMV IgG, determined by the clinical microbiology laboratory, were used for evaluation and optimization. A cutoff optical density (OD) that distinguishes UL144 antibody-positive from antibody-negative sera was established. UL144 A, B, C, and combinations of these antigens were detected in sera. An assay threshold of 0.1 was established and, from a total of 303 sera, the overall sensitivity, specificity, and positive and negative predictive values of the multiplex ELISA were 86.72% (95% confidence interval [CI] 79.59% to 92.07%), 96.57% (92.69% to 98.73%), 94.40% (88.45% to 97.38%), and 91.60% (87.50% to 94.44%), respectively. The inter- and intraassay median coefficients of variation were 0.06 (interquartile range [IQR] 0.56, 0.2) and 0.171 (IQR 0.038, 0.302), respectively. No cross-reactivity was observed with HSV-positive CMV-negative sera. This ELISA gives simple and reproducible results for detection of anti-CMV UL144 IgG. It may assist in differentiating natural infection from CMV vaccines that lack UL144, and may provide an important tool for epidemiological studies of CMV strains.

New cell-signaling pathways for controlling cytomegalovirus replication.

Cytomegalovirus (CMV) is increasingly recognized as an accomplished modulator of cell-signaling pathways, both directly via interaction between viral and cellular proteins, and indirectly by activating metabolic/energy states of infected cells. Viral genes, as well as captured cellular genes, enable CMV to modify these pathways upon binding to cellular receptors, up until generation of virus progeny. Deregulation of cell-signaling pathways appears to be a well-developed tightly balanced virus strategy to achieve the desired consequences in each infected cell type. Importantly and perhaps surprisingly, identification of new signaling pathways in cancer cells positioned CMV as a sophisticated user and abuser of many such pathways, creating opportunities to develop novel therapeutic strategies for inhibiting CMV replication (in addition to standard of care CMV DNA polymerase inhibitors). Advances in genomics and proteomics allow the identification of CMV products interacting with the cellular machinery. Ultimately, clinical implementation of candidate drugs capable of disrupting the delicate balance between CMV and cell-signaling will depend on the specificity and selectivity index of newly identified targets.

Leukocytosis caused by prostaglandin E1 in neonates.

We conducted a retrospective study of neonatal leukocytosis induced by prostaglandin E(1). Among 45 neonates with congenital heart disease, leukocyte counts increased during PGE(1) infusion. We conclude that PGE(1) infusion is a predictable cause of leukocytosis in newborns with congenital heart disease.

Seroprevalence of pertussis antibodies among adolescents in Israel.

BACKGROUND: There is an increasing number of reports of pertussis among older children and adults. The development and licensure of an acellular pertussis vaccine offer the possibility of adult vaccination against the disease. Information on immunity to pertussis in this age group is needed before any vaccination policy can be considered. OBJECTIVES: To study the seroepidemiology of pertussis antibodies in a random sample of adolescents. METHODS: Serum IgG antibodies to whole-cell lysate of Bordetella pertussis were measured by enzyme-linked immunosorbent assay in sera of 533 Israeli military recruits aged 17-18 years. Epidemiologic variables were collected by a questionnaire and analyzed for correlation with pertussis antibodies. RESULTS: Of the sera tested 58.6% were positive for pertussis IgG antibodies, while 35.4% were negative and 6% were borderline. The seropositivity rate was significantly higher among females and non-smokers than among males and smokers. Serum samples of subjects found negative to Bordetella pertussis on recruitment were tested again, using the same ELISA assay, 2-3 years later. Seroconversion during the 3 year military service was detected in 12.5% of 40 subjects. Using the pertussis toxin as the antigen in a subsample of 160 sera, the seroprevalence was lower than that detected by the whole-cell lysate on the same sera (45% vs. 58%). CONCLUSIONS: A significant part of the adolescent population in Israel has low titer of serum IgG antibodies to the multiple antigens of B. pertussis. The relatively low concentration of anti-pertussis antibodies, together with the serological evidence of exposure to the disease indicates that booster immunization with the acellular pertussis vaccine of military recruits should be considered after more information on the incidence of clinical cases of pertussis will be available.

Complications of minocycline therapy for acne vulgaris: case reports and review of the literature.

In order to raise the awareness of the potential side effects that can be induced by minocycline, we report three adolescent girls who experienced serious adverse reactions to this therapy, which was prescribed for acne vulgaris.

Shwachman--Diamond syndrome associated with autoimmune phenomena.

A 2-year-old boy was evaluated for failure to thrive, hypotonia and developmental delay. The child exhibited all the criteria of Shwachman-Diamond syndrome, i.e., short stature, metaphyseal dysostosis, pancreatic insufficiency and neutropenia. Liver function tests were abnormal. Marked edema together with pericardial effusion appeared during the period of follow-up. Hypothyroidism attributed to autoimmune thyroiditis was diagnosed, and other autoantibodies were detected as well. We suggest that an autoimmune baseline profile and follow-up should be part of the work-up and management of patients with Shwachman-Diamond syndrome. Moreover, the finding of autoantibodies might offer a new insight towards understanding the pathogenesis of this condition.

Periodic syndromes of childhood.

Periodic syndromes of childhood comprise a large group of disorders that require long-term follow-up to be diagnosed. Several disorders have fixed rhythmicity and are therefore identified more easily. Other disorders are usually diagnosed after a prolonged follow-up and exclusion of other more common childhood diseases. In general, most of the periodic fever syndromes have a benign prognosis, and their symptomatology tends to improve in the long term. Periodic syndromes without fever or chronic pain syndromes constitute not only a diagnostic dilemma but also a therapeutic challenge. A general diagnostic approach to the periodic syndromes is outlined in Figure 1.

Portal vein thrombosis caused by protein C and protein S deficiency associated with cytomegalovirus infection.

A 4-month-old girl who was examined because of splenomegaly had portal vein thrombosis, which apparently resulted from a combination of cytomegalovirus infection and deficiency of both protein S and protein C. The cytomegalovirus infection, by damaging endothelial cells, may have triggered a cascade of events that was ultimately expressed as portal vein thrombosis resulting from deficiency of proteins S and C.

[Hemophilus influenzae meningitis despite vaccination].

An 18-month-old male previously vaccinated with 4 doses of HbOC developed meningitis caused by Hemophilus influenzae type B. Immunological status was normal and antibody titer to Hemophilus influenzae type b was in the normal range for immunized children. Meningitis due to this organism should be considered even in children who are fully vaccinated.

[Decreased interleukin-1 and interleukin-2 production after splenectomy].

Production of interleukin-1 and of interleukin-2 was measured in 57 splenectomized patients. 11 of them were after elective operations (aged 14-37 years, mean 24) and 46 posttraumatic (aged 20-36, mean 23) and in 20 appropriate controls. There was significant reduction of both interleukins in the splenectomized group, more evident in the elective group. The deficiency was not related to age of patient or time since splenectomy. These results support the view that a consequence of splenectomy is immunoregulatory deficit.

Urinary tract infections with low and high colony counts in young women. Spontaneous remission and single-dose vs multiple-day treatment.

BACKGROUND: The aim of this study was to test whether the spontaneous cure rate is higher in dysuric women with low urinary colony counts (10(2) to 10(4) colony-forming units [cfu] per milliliter) than in women with high colony counts (> or = 10(5) cfu/mL) and whether the response of patients with low colony counts to single-dose treatment is better than those with high colony counts. METHODS: Dysuric women underwent a clinical interview, examination, urine culture, and microscopic examination of urine, and were asked to postpone treatment for 2 days. At that time, the urine tests were repeated. Women with bacterial urinary tract infection were randomized to a group given a single dose of 1200 mg of norfloxacin and to a group treated with 400 mg of norfloxacin twice daily for 7 days. Cure rates were tested at 1 and 5 weeks after treatment. RESULTS: Of 146 women with urinary tract complaints, 113 (25 patients with sterile urine cultures, 21 with low colony counts, and 67 with high colony counts) agreed to postpone antibiotic treatment. Two days after the initial urine culture, only one patient with a low initial colony count had a sterile urine culture (5%, 95% confidence interval, 0% to 14%); and 10 patients (48%, 95% confidence interval, 27% to 69%) had a colony count of 10(5) cfu/mL or more. The rate of spontaneous cure in women with high initial colony count was 7%, 95% confidence interval, 1% to 13%. One week after treatment, the cure rates were 48 (84%) of 57 patients given single-dose treatment vs 49 (98%) of 50 patients treated for 7 days. Five weeks after treatment the rates were 63% and 83%, respectively. The efficacy of single-dose treatment in patients with low urinary counts were similar to those with high counts, and less than that achieved by 7 days of treatment. CONCLUSIONS: The spontaneous cure rate for a 2-day period is minimal in patients with both a low and a high colony count, but half of the patients with low urinary counts will have high colony counts after this interval. The response to single-dose treatment is similar in patients with low and high colony counts, and lower than with multiday treatment.