A polymerase chain reaction for the diagnosis of Haemophilus influenzae type b disease in children and its evaluation during a vaccine trial.

BACKGROUND: Determination of the etiology of pneumonia in young children is difficult because blood culture, the usual method of diagnosis, is positive in only a small proportion of cases. For this reason vaccine trials that include bacterial pneumonia as an endpoint must be large. OBJECTIVES: To determine whether a diagnostic test based on a polymerase chain reaction could be used as an alternative to conventional blood culture for diagnosis of invasive Haemophilus influenzae type b (Hib) infections in young children investigated during the course of a large vaccine trial. METHODS: DNA was extracted from blood culture supernatants and probed for the presence of Hib DNA with a PCR assay with primers derived from the cap gene locus of Hib. Results of the PCR assay were compared with those obtained by conventional culture techniques. RESULTS: Blood cultures were obtained from 1544 children with suspected pneumonia, meningitis or septicemia and from 31 healthy control children who were contacts of cases. Blood culture supernatants were tested for Hib DNA in the PCR test. The sensitivity and specificity of a positive PCR test in blood culture supernatant as against culture of Hib from any normally sterile site were 100 and 99%, respectively. Eleven children had positive Hib PCR tests on blood culture supernatants but were negative by culture. In one of these cases Hib was isolated from a lung aspirate and in two other patients H. influenzae strains other than Hib were obtained from the cerebrospinal fluid. Eight of these 11 children were in the control group. When the results of the PCR assay were used to determine vaccine efficacy, a value of 86% was obtained compared with a figure of 95% obtained when conventional culture techniques were used. CONCLUSIONS: An Hib PCR assay on blood culture supernatants proved to be sensitive and specific for the diagnosis of Hib disease in children. The distribution of PCR-positive, culture-negative cases between Hib-vaccinated and control groups paralleled that of culture-positive cases, suggesting that most of these children had been infected with Hib. A trial of a highly efficacious vaccine provides a novel way for evaluating new diagnostic tests for which there is no standard diagnostic test of 100% reliability.

Epidemiology of invasive pneumococcal disease in the Western Region, The Gambia.

BACKGROUND: Streptococcus pneumoniae is a major cause of morbidity and mortality in young children in the developing world. The recent development of pneumococcal polysaccharide/protein conjugate vaccines may make possible prevention of this infection. However, little is known about the epidemiology of invasive pneumococcal disease in children in the developing world. OBJECTIVES: To determine the incidence and epidemiologic features of invasive pneumococcal disease in children resident in a semiurban area of The Gambia. METHOD: The study was part of a large trial of an Haemophilus influenzae type b vaccine that recruited 42 848 children at the age of 2 months during the period March, 1993, to October, 1995. Follow-up of study children continued until December 31, 1995; therefore the first children to enter the trial were followed for 2.5 years and the last for just a few months. During the period of surveillance, 2256 children were investigated for possible invasive pneumococcal disease when they presented to a hospital or health center. RESULTS: We detected 110 cases of pneumococcal disease. Pneumonia was the most common form of invasive pneumococcal disease observed (75.5% of patients). The incidence of pneumococcal disease was 224 [95% confidence interval (CI) 171, 277] per 100,000 child years among children ages 2 to 11 months, 139 (95% CI 93, 184) per 100,000 among children ages 12 to 23 months and 82 (95% CI 21, 143) per 100,000 among children ages 24 to 35 months. Pneumococci of serogroups 14, 6, 5, 23, 19, 46 and 2 were isolated most frequently. Susceptibility to pneumococcal disease was not increased significantly among Haemophilus influenzae type b-vaccinated children. CONCLUSIONS: The pneumococcus is a major cause of bacterial infection in The Gambia. A proposed nine-valent pneumococcal conjugate vaccine for developing countries containing conjugates of serogroups 1, 4, 5, 6, 9, 14, 18, 19 and 23 would cover 74% of cases of invasive pneumococcal disease in children resident in the Western Region of The Gambia.

Clinical predictors of hypoxaemia in Gambian children with acute lower respiratory tract infection: prospective cohort study.

OBJECTIVES: To determine clinical correlates and outcome of hypoxaemia in children admitted to hospital with an acute lower respiratory tract infection. DESIGN: Prospective cohort study. SETTING: Paediatric wards of the Royal Victoria Hospital and the hospital of the Medical Research Council's hospital in Banjul, the Gambia. SUBJECTS: 1072 of 42 848 children, aged 2 to 33 months, who were enrolled in a randomised trial of a Haemophilus influenzae type b vaccine in the western region of the Gambia, and who were admitted with an acute lower respiratory tract infection to two of three hospitals. MAIN OUTCOME MEASURES: Prevalence of hypoxaemia, defined as an arterial oxygen saturation <90% recorded by pulse oximetry, and the relation between hypoxaemia and aetiological agents. RESULTS: 1072 children aged 2-33 months were enrolled. Sixty three (5.9%) had an arterial oxygen saturation <90%. A logistic regression model showed that cyanosis, a rapid respiratory rate, grunting, head nodding, an absence of a history of fever, and no spontaneous movement during examination were the best independent predictors of hypoxaemia. The presence of an inability to cry, head nodding, or a respiratory rate >/= 90 breaths/min formed the best predictors of hypoxaemia (sensitivity 70%, specificity 79%). Hypoxaemic children were five times more likely to die than non-hypoxaemic children. The presence of malaria parasitaemia had no effect on the prevalence of hypoxaemia or on its association with respiratory rate. CONCLUSION: In children with an acute lower respiratory tract infection, simple physical signs that require minimal expertise to recognise can be used to determine oxygen therapy and to aid in screening for referral. The association between hypoxaemia and death highlights the need for early recognition of the condition and the potential benefit of treatment.

PIP: Acute lower respiratory tract (ALRT) infections cause considerable child morbidity and mortality in developing countries. Oxygen therapy can improve the outcome of children with moderate or severe ALRT infections and, in those with hypoxemia, the severity of hypoxia correlates with outcome. However, since oxygen is not always available in resource-poor countries, rational guidelines must be followed for the use of oxygen and the referral of patients to specialist hospitals. Findings are presented from a prospective cohort study conducted to determine which clinical signs predict hypoxemia and the outcome of hypoxemia among children admitted to hospital with ALRT infection. Findings are based upon the study of 1072 of 42,848 children aged 2-33 months who were enrolled in a randomized trial of a Haemophilus influenzae type b vaccine in western Gambia, and who were admitted with an ALRT infection to 2 of 3 hospitals. 63 (5.9%) had an arterial oxygen saturation level of less than 90%. Logistic regression found cyanosis, a rapid respiratory rate, grunting, head nodding, absence of a history of fever, and no spontaneous movement during examination were significantly associated with hypoxemia. When cyanosis may not be correctly assessed, the inability to cry, head nodding, and a respiratory rate of at least 90 breaths/minute can be useful ways of predicting hypoxemia. Hypoxemic children were 5 times more likely to die than were nonhypoxemic children. The presence of malaria parasitemia had no effect upon the prevalence of hypoxemia or upon its association with respiratory rate.

Comparison of nasal prongs and nasopharyngeal catheter for the delivery of oxygen in children with hypoxemia because of a lower respiratory tract infection.

OBJECTIVE: To determine the best method of oxygen delivery for children in developing countries who have hypoxemia caused by acute lower respiratory tract infection. METHODS: One hundred eighteen children between 7 days and 5 years of age with a lower respiratory tract infection and arterial hemoglobin oxygen saturation (Sao2) less than 90% were randomly selected to receive oxygen by nasopharyngeal (NP) catheter (n = 56) or nasal prongs (n = 62). A crossover study to determine the flow rate necessary to achieve an Sao2 of 95% was performed in 60 children. RESULTS: One hundred twelve children could be oxygenated by the allocated method; in six oxygenation was poor with either method. The mean duration of therapy was 87.5 hours for the prongs and 94.9 hours for the NP catheter (not significant). The median oxygen consumption was 2142 L for prongs and 1692 L for the NP catheter (not significant). In the crossover study the prongs needed, on average, 26% higher oxygen flow rates than the NP catheter to obtain an Sao2 of 95% (p = 0.003). Complete nasal obstruction was observed in 24 of the children (44%) in the NP catheter group and in 8 (13%) in the prongs group (p < 0.001). Eighteen children died, 11 with NP catheter and 7 with prongs (not significant). CONCLUSIONS: Because nasal prongs are less prone to complications, and oxygenation in children is equally effective, they are a more appropriate method than the NP catheter for oxygen delivery to children in developing countries with acute lower respiratory tract infections.

Detection of Haemophilus influenzae and Streptococcus pneumoniae DNA in blood culture by a single PCR assay.

A multiplex PCR assay was developed to screen blood cultures from children in The Gambia with suspected pneumonia for the simultaneous detection of Haemophilus influenzae type b and Streptococcus pneumoniae isolates. Analysis of 295 blood cultures showed that PCR detected the organisms in all samples positive by culture in two samples infected with H. influenzae type b and four samples infected with S. pneumoniae that were culture negative, indicating that this method is sensitive for detecting these organisms in blood cultures.

Randomised trial of Haemophilus influenzae type-b tetanus protein conjugate vaccine [corrected] for prevention of pneumonia and meningitis in Gambian infants.

BACKGROUND: In developing countries, pneumonia and meningitis due to Haemophilus influenzae type b (Hib) are common in children under age 12 months and the mortality from meningitis is high. Protein-polysaccharide conjugate vaccines have brought Hib disease under control in industrialised countries. We did a double-blind randomised trial in The Gambia to assess the efficacy of a Hib conjugate vaccine for the prevention of meningitis, pneumonia, and other invasive diseases due to Hib. METHODS: Between March, 1993, and October, 1995, 42,848 infants were randomly allocated the conjugate vaccine Hib polysaccharide tetanus protein (PRP-T) mixed with diphtheria-tetanus-pertussis vaccine (DTP), or DTP alone at age 2 months, 3 months, and 4 months. Children who presented with signs of invasive Hib were investigated by blood culture and, where appropriate, by lumbar puncture, chest radiograph, or percutaneous lung aspirate. Children were followed up for between 5 and 36 months. FINDINGS: The median ages at which children received the study vaccine were 11 weeks, 18 weeks, and 24 weeks. 83% of children enrolled received all three doses of vaccine. 17 cases of culture-positive Hib pneumonia, 28 of Hib meningitis, and five of other forms of invasive Hib disease were detected amongst the study children. The efficacy of the vaccine for the prevention of all invasive disease after three doses was 95% (PRP-T vaccinees 1, controls 19 [95% CI 67-100]), for the prevention of Hib pneumonia after two or three doses, 100% (vaccinees 0, controls 10 [55-100]), and for the prevention of radiologically defined pneumonia at any time after enrollment, 21.1% (PRP-T vaccinees 198, controls 251 [4.6-34.9]). INTERPRETATION: PRP-T conjugate Hib vaccine prevented most cases of meningitis and pneumonia due to Hib in Gambian infants. The reduction in the overall incidence of radiologically defined pneumonia in PRP-T vaccinees suggests that about 20% of episodes of pneumonia in young Gambian children are due to Hib. The introduction of Hib vaccines into developing countries should substantially reduce childhood mortality due to pneumonia and meningitis.