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R E Huebner

Publications and source records attributed to R E Huebner.

27 records · Page 2Linked to original sources

Current practices in mycobacteriology: results of a survey of state public health laboratories.

Fifty-six state and territorial public health laboratories were surveyed to determine whether currently available rapid methods for the identification and drug susceptibility testing of Mycobacterium tuberculosis were being performed. Forty (71%) laboratories use fluorochrome rather than conventional basic fuchsin stains for screening clinical specimens for acid-fast bacilli. Of the 55 laboratories that routinely culture for mycobacteria, 16 (29%) use the more rapid radiometric methods. Species identification of isolates is done by biochemical tests in 13 (23%) laboratories; 40 (72%) use nucleic acid probes, high-performance liquid chromatography, or the BACTEC p-nitro-alpha-acetylamino-beta-hydroxypropiophenone (NAP) test (rapid tests); 3 laboratories do not perform species identification. Drug susceptibility testing is performed with solid media by 36 of 45 (80%) laboratories, while the more rapid radiometric methods are used by 9 (20%) laboratories. Compared with the laboratories that use conventional methods, laboratories that use rapid methods report results more quickly: for species identification, 43 days (conventional) versus 22 days (rapid); for drug susceptibility testing, 44 days (conventional) versus 31 days (rapid) from specimen processing. Rapid technologies for microscopy and species identification are being used by many, but not all, state and territorial public health laboratories; however, most laboratories do not use the more rapid radiometric methods for routine culture or drug susceptibility testing of mycobacteria. Implementation of such rapid technologies can shorten turnaround times for the laboratory diagnosis of tuberculosis and recognition of drug resistance.

Bacteriological Techniques↗

Usefulness of skin testing with mycobacterial antigens in children with cervical lymphadenopathy.

One hundred twenty-three children with chronic cervical lymphadenopathy were skin-tested with purified protein derivative (PPD)-B (Mycobacterium intracellulare), PPD-Y (Mycobacterium kansasii), PPD-G (Mycobacterium scrofulaceum) (nontuberculous mycobacterial antigens (NTMags)) and PPD-T (Mycobacterium tuberculosis). Children with culture-confirmed mycobacterial disease had significantly larger reactions to NTMags and were 6 times more likely to have PPD-B responses of greater than or equal to 10 mm than those with negative microscopy/culture results. Children with acid-fast bacilli present in clinical specimens but with negative culture results were 3 times more likely to have greater than or equal to 10 mm induration to PPD-B than those with negative microscopy/culture results. In all groups except those with culture-confirmed M. tuberculosis, responses to PPD-T were significantly smaller than those to the NTMags. We conclude that NTMags, particularly PPD-B, may be useful in diagnosing childhood mycobacterial cervical adenopathy; however, their usefulness in distinguishing disease caused by M. tuberculosis from that resulting from other mycobacteria is unknown.

Adolescent↗

Evaluation of the clinical usefulness of mycobacterial skin test antigens in adults with pulmonary mycobacterioses.

A double-blind, multicenter study was conducted to evaluate the usefulness of mycobacterial skin test antigens for the specific diagnosis of adult pulmonary mycobacterial disease. The skin test antigens used were PPD-T (M. bovis) and PPD-B (M. intracellulare), made bioequivalent to 5 TU PPD-S through bioassay in human subjects. Of the 192 adults (18 yr of age or older), those with disease caused by M. tuberculosis (MTB) had significantly larger reactions to PPD-T than did those with disease caused by nontuberculous mycobacteria (NTM) or those with negative culture results (NEG)(13.41 mm versus 4.87 and 4.96 mm, respectively, p less than 0.001). The mean induration to PPD-B in NTM was not different from that in MTB or NEG. Defining a "positive" to be greater than or equal to 10 mm induration and a size difference of greater than or equal to 3 mm between PPD-T and PPD-B, the sensitivity, specificity, and positive predictive value (PPV) for PPD-T in diagnosing MTB versus NTM was 29, 90, and 75%. Corresponding values for PPD-B and NTM disease were 70, 61, and 64%. Dual testing was less useful in distinguishing disease caused by any of the mycobacteria from NEG. Although the sensitivity of PPD-B, made bioequivalent to PPD-S, was high, the specificity and PPV were low. We conclude that this preparation of PPD-B is no more useful in distinguishing adult pulmonary disease caused by NTM than is PPD-T alone.

Double-Blind Method↗

Immunomodulation and Chlamydia: immunosuppression and the protective immune response to C. psittaci in mice.

Mice immunized intramuscularly with a low dose, viable inoculum of C. psittaci survived an otherwise lethal intraperitoneal challenge with the homologous chlamydial strain. Immunized animals were not protected from intraperitoneal challenge by the unrelated pathogen, Listeria monocytogenes. Spleen cells from animals that exhibited protective immunity were suppressed in their proliferative responses to mitogens or chlamydial antigen in an in vitro blastogenic assay. This suppression was transferable to normal spleen cells by adding irradiated cells from immunized animals to normal cell populations. The degree of normal cell blastogenic suppression was dependent on the ratio of irradiated immune to normal cells present in the assay medium. Suppression of humoral responses was demonstrated in vivo. Immunized animals were incapable of producing antibody secreting cells to sheep red blood cells after an intraperitoneal inoculation of SRBC. Unimmunized animals produced a significant number of plaque forming cells as measured by a direct plaque forming cell assay. Lymphokine activity was not impaired in spleen cells from mice that exhibited other manifestations of suppression. Taken together, these data provide evidence to indicate that the induction of suppression may not correlate with increased pathogenesis, but rather be closely associated with protective immunity. Data also provide circumstantial evidence to indicate that lymphokine induction may be important in the development of protective immunity to C. psittaci in the mouse.

Animals↗

In vivo-activated mononuclear phagocytes and protective immunity to chlamydiae in mice.

Peritoneal macrophages (M phi s) collected from Chlamydia psittaci 6BC-immune mice after intraperitoneal challenge with 10(6) 6BC (immune-boosted [IB] M phi s) were compared by various functional criteria with other in vivo- and in vitro-activated M phi populations. While casein-, protease peptone-, and thioglycolate (Thio)-elicited M phi s were equally susceptible to in vitro infection with 6BC, IB M phi s did not support chlamydial growth and M phi s from Mycobacterium tuberculosis BCG- or Listeria monocytogenes-sensitized mice exhibited intermediate susceptibility to infection. The resistance of IB M phi s was not due to the ingestion of fewer 6BC organisms, nor were these cells persistently infected, since chlamydiae could not be recovered from infected IB M phi s after in vitro infection, even after extended incubation times. In contrast, Thio M phi s stimulated in vitro with gamma interferon (IFN-gamma), with or without lipopolysaccharide, resulted in cells that exhibited chlamydiastatic activity which was lost shortly after IFN-gamma was removed from the culture medium. Conversely, the antichlamydial activity of IB M phi s was stable over time but not through the production of autostimulatory cytokines, as evidenced by the lack of stimulation of Thio M phi s to restrict 6BC replication in coculture experiments. IB M phi s exhibited enhanced oxidative activity, but anti-IFN-gamma antibody did not abrogate this response. IB M phi s were recovered only from immunized mice that survived an otherwise lethal 6BC intraperitoneal challenge. These cells appear to be important for development of protective immunity to chlamydiae, and evidence suggests that stimulation by cytokines other than IFN-gamma (with or without lipopolysaccharide) is required for the observed heightened in vivo activation.

Animals↗

Nasopharyngeal carriage and antimicrobial resistance in isolates of Streptococcus pneumoniae and Haemophilus influenzae type b in children under 5 years of age in Botswana.

OBJECTIVES: A prospective survey was conducted to determine the prevalence of asymptomatic nasopharyngeal carriage of Streptococcus pneumoniae and Haemophilus influenzae type b in children under 5 years of age in Botswana and to determine the antibiotic resistance patterns of these organisms to commonly used antimicrobial agents. METHODS: Children 2 months to 5 years of age (n = 249) were recruited from outpatient clinics in Gaborone and Francistown, and 29 were sampled from the pediatric wards at Princess Marina (Gaborone) and Nyangabgwe (Francistown) Hospitals. Nasopharyngeal specimens were collected and the carriage and antibiotic resistance of S. pneumoniae and H. influenzae type b were determined. Analyses of risk factors associated with carriage and resistance were performed. RESULTS: Streptococcus pneumoniae was isolated from 69% of the outpatient children in Gaborone and 85% of the children in Francistown; the carriage rate in hospitalized children was 36% and 33% in Gaborone and Francistown, respectively. Approximately half of the isolates at both sites were resistant to at least one antibiotic, the most common being cotrimoxazole and penicillin. Resistance to three or more antibiotics (multiple resistance) was found in less than 10% of the isolates. Most penicillin resistance at both sites was at the intermediate level; however, almost 20% of the isolates demonstrated high-level resistance to cotrimoxazole. The most prevalent serogroups or serotypes of antibiotic-resistant isolates were 14, 19F, 19A, 6A, 6B, and 4. No risk factors for antibiotic resistance were identified. Haemophilus influenzae type b was isolated from 8% of the children in Gaborone and from 3% of the children in Francistown. Almost a third of the isolates were resistant to ampicillin. CONCLUSIONS: The high levels of antibiotic resistance in pneumococci isolated from children in Botswana suggest that the clinical management of meningitis and otitis media with a b-lactam antibiotic may fail in a significant proportion of cases and that empiric first-line use of cefotaxime or ceftriaxone for meningitis and higher dose amoxicillin (90 mg/kg/day) for otitis media is recommended. The levels of penicillin resistance in this study would not impact on the management of pneumonia with amoxicillin.

Anti-Bacterial Agents↗