Culture-Negative Neonatal Meningitis and Endocarditis Caused by Streptococcus agalactiae

Abstract

We describe a case of culture-negative meningitis and endocarditis caused by Streptococcus agalactiae in a 27-day-old boy. S. agalactiae was detected in cerebrospinal fluid and serum by broad-spectrum PCR amplification. CASE REPORTA previously healthy neonate aged 27 days was admitted to the pediatric department of Robert Ballanger Hospital (Aulnay sous bois, France) because of fever onset the day before. On initial evaluation he was pale and continuously crying, and he had a temperature of 40.8°C. Neurologic examination was normal, except for an anterior bulging fontanelle. Cardiologic examination showed a 2-3/6 systolic murmur at the apex. The ears, lungs, and abdomen appeared normal. A chest radiograph was normal. Blood, cerebrospinal fluid (CSF), and urine samples were immediately taken for culture. Laboratory findings included a white blood cell (WBC) count of 21,000/ mm 3 (44% polymorphonuclear neutrophils) and a C-reactive protein (CRP) level of 157 mg/liter. Urine laboratory examination was normal. Lumbar puncture yielded cloudy CSF with 50 red blood cells/mm 3 and 7,000 WBC/mm 3 (89% polymorphonuclear neutrophils), an elevated protein level (1.75 g/liter), and a low glucose concentration (1.4 mmol/liter; serum concentration, 3.6 mmol/liter). Direct examination with Gram staining was negative. Tests for Streptococcus agalactiae and Escherichia coli K1 soluble antigens using the Pastorex Meningitis kit (Bio-Rad, Marne-La-Coquette, France) were negative in urine and CSF.Empirical intravenous antibacterial chemotherapy for neonatal meningitis was immediately started, with cefotaxime (200 mg/kg of body weight/24 h), amoxicillin (200 mg/kg/24 h), and amikacin (15 mg/kg/24 h). Blood and CSF cultures remained sterile after 5 days. A second lumbar puncture performed 48 h after admission yielded sterile bloody CSF. The temperature and CRP normalized after 2 and 5 days of treatment, respectively. Amikacin and amoxicillin were withdrawn, and cefotaxime (200 mg/kg/24 h) was continued. The boy's clinical status improved, and neurological examination remained normal. Because the systolic murmur was still present at 15 days of hospitalization, the infant was transferred to the neonatal cardiology unit of Robert-Debré Hospital for further investigations. An echocardiography showed grade 2/4 mitral regurgitation and a 3 mm vegetation on the mitral valve. The other valves were normal.Because of sterile cultures, an admission CSF sample and a day 1 serum sample, both stored at Ϫ20°C, were sent to Robert-Debré Hospital for molecular diagnosis by universal 16S rRNA PCR amplification. DNA was extracted with the QIAmp DNA Mini kit (QIAGEN, Courtaboeuf, France). The initial PCR mixture (final volume, 50 l) contained 0.4 M each universal primer, 200 M each deoxynucleoside triphosphate, and 1 U of HotStarTaq DNA polymerase (QIAGEN) in 1ϫ amplification buffer supplied by the manufacturer; the final concentration of MgCl 2 was 3 mM. The universal primers 779F (5Ј-CAAACAGGATTAGATACCC T-3Ј) and 1191R (5Ј-CGTCATCCCCACCTTCCTCC-3Ј) corr...

Implementation of the FilmArray ME panel in laboratory routine using a simple sample selection strategy for diagnosis of meningitis and encephalitis

“…One sample positive for S. agalactiae in FilmArray E panel showed no inhibition of growth in B. subtilis inhibition assay. Notably, culture negative yet PCR positive infectious meningitis cases caused by S. agalactiae have been described previously, underlining the importance of molecular diagnostic for optimization of patient management [40][41][42]. Otherwise, it cannot be excluded that in our sample, FilmArray ME detection of S.agalactiae was indeed false positive and the communication of the test result might have led to a dispensable antibiotic administration.…”

New Real-Time PCR-Based Method for Kingella kingae DNA Detection: Application to Samples Collected from 89 Children with Acute Arthritis

“…Depending on the sites of infection and the pathogens, DNA may persist for several weeks (3,15,18) or less than 48 h (17,22). To our knowledge only one study has investigated this property in cases of septic arthritis (23).…”

Persistent Bloodstream Infection with Kocuria rhizophila Related to a Damaged Central Catheter

“…In order to clarify this discordant test and confirm K. varians, we used 16S rRNA gene analysis. DNA extraction and 16S rRNA gene sequence analysis were performed as previously described (16). The procedure was performed on 11 clinical isolates: four recovered in 2006, six in 2008, and one in 2009.…”