SUMMARY Kingella kingae is a common etiology of pediatric bacteremia and the leading agent of osteomyelitis and septic arthritis in children aged 6 to 36 months. This Gram-negative bacterium is carried asymptomatically in the oropharynx and disseminates by close interpersonal contact. The colonized epithelium is the source of bloodstream invasion and dissemination to distant sites, and certain clones show significant association with bacteremia, osteoarthritis, or endocarditis. Kingella kingae produces an RTX (repeat-in-toxin) toxin with broad-spectrum cytotoxicity that probably facilitates mucosal colonization and persistence of the organism in the bloodstream and deep body tissues. With the exception of patients with endocardial involvement, children with K. kingae diseases often show only mild symptoms and signs, necessitating clinical acumen. The isolation of K. kingae on routine solid media is suboptimal, and detection of the bacterium is significantly improved by inoculating exudates into blood culture bottles and the use of PCR-based assays. The organism is generally susceptible to antibiotics that are administered to young patients with joint and bone infections. β-Lactamase production is clonal, and the local prevalence of β-lactamase-producing strains is variable. If adequately and promptly treated, invasive K. kingae infections with no endocardial involvement usually run a benign clinical course.

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A double-blind, randomized study involving 264 toddlers attending day care centers was conducted to document the effect of a 9-valent pneumococcal conjugate vaccine on the carriage rate of pneumococci. Of 3750 cultures done on nasopharyngeal samples obtained from subjects during a 2-year follow-up period after vaccination, 65% were positive for Streptococcus pneumoniae. In all age windows, the rate of carriage of vaccine-type pneumococci was lower among subjects who received the pneumococcal vaccine than among control subjects, because the acquisition rate was lower in the former group. The effect was most pronounced among subjects aged < or =36 months. The sample size enabled us to study protection against carriage of S. pneumoniae serotypes 6B, 9V, 14, 19F, and 23F; significant protection against all serotypes except 19F was seen in the pneumococcal-vaccine group. The rate of carriage of serotype 6A (not included in the vaccine) was also reduced significantly, but the rate of carriage of serotype 19A (not included in the vaccine) was not. The rate of carriage of non-vaccine-type pneumococci (excluding serotype 6A) was higher in the pneumococcal-vaccine group than in the control group.

Children 12-18 months old were randomized to receive one dose of a conjugate heptavalent pneumococcal vaccine, two doses of the same vaccine, or one dose of a 23-valent native polysaccharide vaccine. Before immunization, pneumococci included in the conjugate vaccine were isolated from 24% of the children, and an antibiotic-resistant pneumococcus was isolated from 22% of the children. The vaccines had no effect on carriage of non-vaccine-type pneumococci. In contrast, there was a significant reduction in carriage of vaccine-type pneumococci 3 months after one dose and 1 month after a second dose of conjugate vaccine (from 25% to 9% and 7%, respectively; P < .001). No effect was seen after vaccination with the nonconjugate vaccine. One year after immunization, carriage of antibiotic-resistant vaccine-type pneumococci in children receiving conjugate vaccine was lower than that in children receiving the nonconjugate vaccine (4% vs. 14%, P = .042). Conjugate pneumococcal vaccines may reduce spread of pneumococci in the community.

For years, quantitative blood cultures found only limited use as aids in the diagnosis and management of septic patients because the available methods were cumbersome, labor intensive, and practical only for relatively small volumes of blood. The development and subsequent commercial availability of lysis-centrifugation direct plating methods for blood cultures have addressed many of the shortcomings of the older methods. The lysis-centrifugation method has demonstrated good performance relative to broth-based blood culture methods. As a result, quantitative blood cultures have found widespread use in clinical microbiology laboratories. Most episodes of clinical significant bacteremia in adults are characterized by low numbers of bacteria per milliliter of blood. In children, the magnitude of bacteremia is generally much higher, with the highest numbers of bacteria found in the blood of septic neonates. The magnitude of bacteremia correlates with the severity of disease in children and with mortality rates in adults, but other factors play more important roles in determining the patient's outcome. Serial quantitative blood cultures have been used to monitor the in vivo efficacy of antibiotic therapy in patients with slowly resolving sepsis, such as disseminated Mycobacterium avium-M. intracellulare complex infections. Quantitative blood culture methods were used in early studies of bacterial endocarditis, and the results significantly contributed to our understanding of the pathophysiology of this disease. Comparison of paired quantitative blood cultures obtained from a peripheral vein and the central venous catheter has been used to help identify patients with catheter-related sepsis and is the only method that does not require removal of the catheter to establish the diagnosis. Quantitation of bacteria in the blood can also help distinguish contaminated from truly positive blood cultures; however, no quantitative criteria can invariably differentiate contamination from bacteremia.

Kingella kingae is being recognized increasingly as a common etiology of pediatric osteoarticular infections, bacteremia, and endocarditis, which reflects improved culture methods and use of nucleic acid-amplification techniques in clinical microbiology laboratories. K kingae colonizes the posterior pharynx of young children and is transmitted from child to child through close personal contact. Day care attendance increases the risk for colonization and transmission, and clusters of K kingae infections among day care center attendees have been reported. Key virulence factors in K kingae include type IV pili and a potent RTX toxin. In previously healthy children, >95% of K kingae infections are diagnosed between the ages of 6 and 48 months. Among children with underlying medical conditions, K kingae disease may occur at older ages as well. The clinical presentation of K kingae disease is often subtle and may be associated with normal levels of acute-phase reactants, which underscores the importance of a high index of suspicion. K kingae is usually susceptible to ß-lactam antibiotics, and infections typically respond well to medical treatment, with the exception of cases of endocarditis.

Field studies of Streptococcus pneumoniae (pneumococci) nasopharyngeal (NP) colonization are hampered by the need to directly plate specimens in order to ensure isolate viability. A medium containing skim milk, tryptone, glucose, and glycerin (STGG) has been used to transport and store NP material, but its ability to preserve pneumococci has not been evaluated. Our objective was to qualitatively and semiquantitatively evaluate the ability of STGG to preserve pneumococci in NP secretions. Entwined duplicate calcium alginate NP swab samples were obtained from children. One swab was plated directly onto a gentamicin blood agar plate; the other was placed in STGG. Growth from the directly plated specimen was compared with growth from an STGG aliquot immediately cultured or stored at ؊70°C for 9 weeks, ؊20°C for 9 weeks, or 4°C for 5 days. Of 186 specimens, 96 (52%) were positive for pneumococci from the direct plating; 94 (98%) of these were positive from the fresh STGG specimen. Pneumococci were recovered from all 38 positive specimens frozen at ؊70°C, all 18 positive specimens frozen at ؊20°C, and 18 of 20 positive specimens stored at 4°C. Recovery of pneumococci after storage of NP material in STGG medium at ؊70°C is at least as good as that from direct plating. Storage at ؊20°C is also acceptable. Storage at 4°C for 5 days is not ideal.Streptococcus pneumoniae (pneumococci) is the most important cause of bacterial otitis media, pneumonia, bacteremia, and meningitis among children worldwide (12,15,17). Pneumococci are also important because the rate of nonsusceptibility to various classes of antimicrobial agents, such as penicillins and cephalosporins, is rising throughout the United States and worldwide (4,18,19). Prevention efforts have been hampered by the lack of a vaccine which is immunogenic for important serotypes in children younger than 2 years of age. A seven-valence pneumococcal conjugate vaccine (Prevnar; Wyeth Lederle Vaccines) which is immunogenic and efficacious in this age group recently has been licensed in the United States for use among children through 9 years of age and is recommended routinely for those under 2 years of age (1, 3, 16). The effect of this and other conjugate pneumococcal vaccines on nasopharyngeal (NP) colonization is a subject of intense investigation.It is well known that pneumococci are spread from person to person via the respiratory route. NP colonization studies have shown that people acquire pneumococci at a young age, carry these organisms for various periods of time, may carry more than one serotype at a time, and transmit these organisms to others with whom they are in close contact (2,5,(9)(10)(11)14). Many studies of the dynamics and ecology of pneumococcal NP carriage, particularly in the setting of new conjugate pneumococcal vaccines, will be performed in settings where microbiologic facilities are not readily available.An optimal medium has not been validated for the transport, preservation, and recovery of pneumococci from NP material. One medium, STGG (skim m...

SUMMARY The clinical presentation of brucellosis in humans is variable and unspecific, and thus, laboratory corroboration of the diagnosis is essential for the patient’s proper treatment. The diagnosis of brucellar infections can be made by culture, serological tests, and nucleic acid amplification assays. Modern automated blood culture systems enable detection of acute cases of brucellosis within the routine 5- to 7-day incubation protocol employed in clinical microbiology laboratories, although a longer incubation and performance of blind subcultures may be needed for protracted cases. Serological tests, though they lack specificity and provide results that may be difficult to interpret in individuals repeatedly exposed to Brucella organisms, nevertheless remain a diagnostic cornerstone in resource-poor countries. Nucleic acid amplification assays combine exquisite sensitivity, specificity, and safety and enable rapid diagnosis of the disease. However, long-term persistence of positive molecular test results in patients that have apparently fully recovered is common and has unclear clinical significance and therapeutic implications. Therefore, as long as there are no sufficiently validated commercial tests or studies that demonstrate an adequate interlaboratory reproducibility of the different homemade PCR assays, cultures and serological methods will remain the primary tools for the diagnosis and posttherapeutic follow-up of human brucellosis.