We describe an outbreak of necrotizing enterocolitis (NEC) that occurred in the neonatal intensive care unit of our hospital. A total of 12 neonates developed NEC in June-July 1998. For two of them, twin brothers, the NEC turned out to be fatal. Enterobacter sakazakii, a known contaminant of powdered milk formula, was isolated from a stomach aspirate, anal swab, and/or blood sample for 6 of the 12 neonates. A review of feeding procedures revealed that 10 of the 12 patients were fed orally with the same brand of powdered milk formula. E. sakazakii was isolated from the implicated prepared formula milk as well as from several unopened cans of a single batch. Molecular typing by arbitrarily primed PCR (AP-PCR) confirmed, although partially, strain similarity between milk and patient isolates. No further cases of NEC were observed after the use of the contaminated milk formula was stopped. With this outbreak we show that intrinsic microbiological contamination of powdered milk formula can be a possible contributive factor in the development of NEC, a condition encountered almost exclusively in formula-fed premature infants. The use of sterilized liquid milk formula in neonatal care could prevent problems with intrinsic and extrinsic contamination of powdered milk formula.
The LIAISON system is a fully automated system based on chemiluminescence and antigen bound to magnetic microparticles. The system allows fast and precise measurement of Toxoplasma-specific immunoglobulin G (IgG) and IgM antibody levels and measurement of the IgG avidity index even at low levels of Toxoplasma-specific IgG antibodies in a single step without manual interference. Seven European centers participated in a multicenter evaluation of the LIAISON system. The sensitivity and specificity of the LIAISON system compared to the Sabin-Feldman dye test were 99.3 and 96.8%, respectively. In a comparison of the LIAISON Toxoplasma-specific IgM assay with an immunosorbent agglutination assay, the LIAISON assay had a sensitivity of 96.7% and a specificity of 95.4%. The LIAISON IgG assay showed agreements of 91, 100, and 100% with the AXSYM IgG (Abbott), VIDAS IgG (bioMérieux), and Platelia IgG (Bio-Rad) assays, respectively. The LIAISON IgM assay showed agreements of 95% with the AXSYM IgM and Platelia IgM assays, 96% with the ISAGA IgM assay (bioMérieux), and 97% with the VIDAS IgM assay. The coefficient of correlation between the LIAISON system and the VIDAS Toxoplasma-specific IgG avidity index was 0.81. By use of the Toxoplasma-specific IgG avidity index assay with specific IgM-positive samples, the diagnosis of infection with Toxoplasma gondii in early pregnancy has been improved significantly. The LIAISON avidity assay is a valuable assay for the exclusion of recently acquired infection with T. gondii (less than 4 months) in pregnant women, and it decreases significantly the necessity for follow-up testing.Primary maternal infection with Toxoplasma gondii carries the risk of transmitting the infection to the fetus, resulting in congenital infection. Congenital infection of the fetus in women infected just before conception is extremely rare, and even during the first few weeks of pregnancy, the maternalfetal transmission rate is low (6). It is therefore essential to estimate the time of infection as precisely as possible to properly manage the risk to the fetus of a maternal infection. Low levels of Toxoplasma-specific immunoglobulin M (IgM) antibodies may be found for up to several years, and the mere demonstration of low levels of Toxoplasma-specific IgM antibodies is therefore not a sign by itself of acute infection with T. gondii (16,17). The measurement of the avidity of IgG antibodies for T. gondii infections was first demonstrated in 1989 (8, 9, 15) and since then has been further developed (4,18).A study of the diagnostic value of various tests for acute infections with T. gondii, including Toxoplasma-specific IgG, IgM, and IgA antibodies and the IgG avidity index, showed that the combination of a sensitive test for Toxoplasma-specific IgM antibodies and a Toxoplasma-specific IgG avidity assay had the highest predictive value with regard to the time of infection (23).Since previous studies showed that some individuals have low-avidity IgG antibodies many months after infection, we also tested the hypo...
Two protocols for the extraction of cytomegalovirus (CMV) DNA and two methods for the amplification of CMV DNA in dried blood spots were evaluated for the retrospective diagnosis of congenital CMV infection. During the period from 1996 to 2006, a urine screening program detected 76 congenitally infected neonates. Stored Guthrie cards with blood from 55 cases and 12 controls were tested. Two spots of dried blood were cut from each card and evaluated in two centers. CMV DNA was extracted from a whole single spot. Center 1 used phenol-chloroform extraction and ethanol precipitation followed by a conventional PCR. Center 2 used the NucliSens easyMAG automated DNA/RNA extraction platform (bioMérieux) followed by a real-time PCR. For evaluation of the extraction method, DNA extracted from each blood spot was evaluated by the amplification method used by the collaborating center. The sensitivities were 66% for center 1 and 73% for center 2. None of the controls were positive. A sensitivity as high as 82% could be obtained by combining the most sensitive extraction method (the phenol-chloroform procedure) with the most sensitive PCR method (real-time PCR). The detection rate was not influenced by the duration of storage of the spots. The sensitivity was higher with blood from congenitally infected cases due to a primary maternal CMV infection, regardless of the protocol used. However, the difference reached significance only for the least-sensitive protocol (P ؍ 0.036).Cytomegalovirus (CMV) infection during pregnancy can be transmitted to the fetus, resulting in a congenital infection. Congenital CMV infection (cCMV) occurs in 0.2 to 2.2% of live-born infants. An incidence of 0.62% is found in the Brussels, Belgium, region (11).The standard diagnosis of cCMV relies on the isolation of the virus from urine or saliva collected in the first 2 weeks of life. The diagnosis of cCMV infection in children older than 2 weeks cannot be made or excluded on the basis of viral isolation. Blood is routinely collected from neonates in the first 2 weeks of life and is stored as dried blood spots (DBSs) on Guthrie cards. The detection of CMV DNA on these stored DBSs could be an opportunity to diagnose cCMV during later life, when symptoms suggestive of cCMV develop (3).The aim of this study was to test the clinical sensitivity of CMV DNA PCR with DBSs from consecutive cases of neonates with cCMV infections. We evaluated two methods for the extraction of CMV DNA and two methods for the amplification of CMV DNA from DBSs on Guthrie cards for the retrospective diagnosis of cCMV. MATERIALS AND METHODSThe study protocol was approved by the Committee of Medical Ethics of the UZ Brussel. cCMV infections. From 1996 to 2006 a neonatal screening program (11) detected cCMV in 76 newborns at the Universitair Ziekenhuis Brussels. The diagnosis of cCMV infection was made by isolation of the virus from urine within 7 days of birth. Blood from the first consecutive 57 cases and from the last 4 cases was retrieved and placed on Guthrie cards (no. 903 ...
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