Performance of spirometry assessment at TB diagnosis.

BACKGROUND: Spirometry is considered relevant for the diagnosis and monitoring of post-TB lung disease. However, spirometry is rarely done in newly diagnosed TB patients.METHODS: Newly diagnosed, microbiologically confirmed TB patients were recruited for the study. Spirometry was performed within 21 days of TB treatment initiation according to American Thoracic Society/European Respiratory Society guidelines. Spirometry analysis was done using Global Lung Initiative equations for standardisation.RESULTS: Of 1,430 eligible study participants, 24.7% (353/1,430) had no spirometry performed mainly due to contraindications and 23.0% (329/1,430) had invalid results; 52.3% (748/1,430) of participants had a valid result, 82.8% (619/748) of whom had abnormal spirometry. Of participants with abnormal spirometry, 70% (436/619) had low forced vital capacity (FVC), 6.1% (38/619) had a low ratio of forced expiratory volume in 1 sec (FEV1) to FVC, and 19.1% (118/619) had low FVC, as well as low FEV1/FVC ratio. Among those with abnormal spirometry, 26.3% (163/619) had severe lung impairment.CONCLUSIONS: In this population, a high proportion of not performed and invalid spirometry assessments was observed; this was addressed by removing tachycardia as a (relative) contraindication from the study guidance and retraining. The high proportion of patients with severe pulmonary impairment at the time of TB diagnosis suggests a huge morbidity burden and calls for further longitudinal studies on the relevance of spirometry in predicting chronic lung impairment after TB.

A phase IIb, open-label, randomized controlled dose ranging multi-centre trial to evaluate the safety, tolerability, pharmacokinetics and exposure-response relationship of different doses of delpazolid in combination with bedaquiline delamanid moxifloxacin in adult subjects with newly diagnosed, uncomplicated, smear-positive, drug-sensitive pulmonary tuberculosis.

BACKGROUND: Linezolid is an effective, but toxic anti-tuberculosis drug that is currently recommended for the treatment of drug-resistant tuberculosis. Improved oxazolidinones should have a better safety profile, while preserving efficacy. Delpazolid is a novel oxazolidinone developed by LegoChem Biosciences Inc. that has been evaluated up to phase 2a clinical trials. Since oxazolidinone toxicity can occur late in treatment, LegoChem Biosciences Inc. and the PanACEA Consortium designed DECODE to be an innovative dose-ranging study with long-term follow-up for determining the exposure-response and exposure-toxicity relationship of delpazolid to support dose selection for later studies. Delpazolid is administered in combination with bedaquiline, delamanid and moxifloxacin. METHODS: Seventy-five participants with drug-sensitive, pulmonary tuberculosis will receive bedaquiline, delamanid and moxifloxacin, and will be randomized to delpazolid dosages of 0 mg, 400 mg, 800 mg, 1200 mg once daily, or 800 mg twice daily, for 16 weeks. The primary efficacy endpoint will be the rate of decline of bacterial load on treatment, measured by MGIT liquid culture time to detection from weekly sputum cultures. The primary safety endpoint will be the proportion of oxazolidinone class toxicities; neuropathy, myelosuppression, or tyramine pressor response. Participants who convert to negative liquid media culture by week 8 will stop treatment after the end of their 16-week course and will be observed for relapse until week 52. Participants who do not convert to negative culture will receive continuation phase treatment with rifampicin and isoniazid to complete a six-month treatment course. DISCUSSION: DECODE is an innovative dose-finding trial, designed to support exposure-response modelling for safe and effective dose selection. The trial design allows assessment of occurrence of late toxicities as observed with linezolid, which is necessary in clinical evaluation of novel oxazolidinones. The primary efficacy endpoint is the change in bacterial load, an endpoint conventionally used in shorter dose-finding trials. Long-term follow-up after shortened treatment is possible through a safety rule excluding slow-and non-responders from potentially poorly performing dosages. TRIAL REGISTRATION: DECODE was registered in ClinicalTrials.gov before recruitment start on 22 October 2021 (NCT04550832).

Diagnosis of paediatric TB using Xpert® MTB/RIF Ultra on fresh respiratory samples.

OBJECTIVE: To evaluate the diagnostic accuracy of Xpert® MTB/RIF Ultra (Ultra) on fresh respiratory samples for the diagnosis of pulmonary TB (PTB) in children.METHODS: Between July 2017 and December 2019, children with presumed TB were prospectively enrolled at clinical sites in three African countries. Children were assessed using history, physical examination and chest X-ray. Sputum or gastric aspirate samples were analysed using Ultra and culture. The diagnostic accuracy of Ultra was calculated against culture as the reference standard.RESULTS: In total, 547children were included. The median age was 4.7 years, 77 (14.1%) were HIV infected and 77 (14.1%) had bacteriologically confirmed TB. Ultra detected an additional 20 cases in the group of children with negative culture results. The sensitivity of Ultra was 66.3% (95% CI 47-82), and the specificity was 95.4% (95% CI 89-99) when assessed against culture as the reference standard.CONCLUSION: Despite the improved performance of Ultra as compared to Xpert as was previously reported, its sensitivity remains sub-optimal for the detection of TB in children. Ultra detected additional 20 cases which otherwise could not have been detected by culture alone, suggesting that the latter is an imperfect reference standard.

Connecting the Dots from Professional Development to Student Learning.

Following professional development (PD), implementation of contemporary topics into high school biology requires teachers to make critical decisions regarding integration of novel content into existing course scope and sequence. Often exciting topics, such as neuroscience, do not perfectly align with standards. Despite commitment to enacting what was learned in the PD, teachers must adapt novel content to their perceptions of good teaching, local context, prior knowledge of their students, and state and district expectations. How teachers decide to integrate curricula encountered from PD programs may affect student outcomes. This mixed-methods study examined the relationship between curricular application strategies following an inquiry-based neuroscience PD and student learning. Post-PD curricular implementation was measured qualitatively through analysis of teacher action plans and classroom observations and quantitatively using hierarchical linear modeling to determine the impact of implementation on student performance. Participation in neuroscience PD predicted improved student learning compared with control teachers. Of the two distinct curricular implementation strategies, enacting a full unit produced significantly greater student learning than integrating neuroscience activities into existing biology units. Insights from this analysis should inform teacher implementation of new curricula after PD on other contemporary biology topics.

[Temporary hyposmia in COVID-19 patients]. / Temporäre Hyposmie bei COVID-19-Patienten.

BACKGROUND: This is a report on the high incidence of olfactory dysfunction in COVID-19 patients in the first cohort of COVID-19 patients in Germany (Webasto cluster). METHODS: Loss of sense of smell and/or taste was reported by 26 of 63 COVID-19 patients (41%), whereas only 31% of the patients experiencing hyposmia had simultaneous symptoms of rhinitis. Smell tests were performed in 14 of these patients and taste tests in 10. The measurements were conducted in a patient care setting in an early COVID-19 cohort. RESULTS: An olfactory disorder was present in 10/14 patients, before as well as after nasal decongestion. In 2 of these patients, hyposmia was the leading or only symptom of SARS-CoV­2 infection. All tested patients reported recovery of smell and/or taste within 8 to 23 days. CONCLUSION: The data imply that a) COVID-19 can lead to hyposmia in a relevant number of patients, the incidence was approximately 30% in this cohort; b) in most cases, the olfactory disturbance was not associated with nasal obstruction, thus indicating a possible neurogenic origin; and c) the olfactory disorder largely resolved within 1-3 weeks after the onset of COVID-19 symptoms. There were no indications of an increased incidence of dysgeusia. These early data may help in the interpretation of COVID-19-associated hyposmia as well as in the counseling of patients, given the temporary nature of hyposmia observed in this study. Furthermore, according to the current experience, hyposmia without rhinitic obstruction can be the leading or even the only symptom of a SARS-CoV­2 infection.

Identifying Dormant Growth State of Mycobacteria by Orthogonal Analytical Approaches on a Single Cell and Ensemble Basis.

Tuberculosis is currently the single most deadly infectious disease in the world and a public health priority as defined by WHO. Although the disease is in general curable, treatment success is hampered by the necessity of a long and side effect prone treatment. Low treatment efficiency may be partly due to the special growth states that mycobacteria enter to avoid being killed by antibiotics and to persist longer within the host. Such growth states have been recently defined as dormant or persistent. We produced dormant model-organism cultures using an acidification model and characterized those by a multilayered approach using mass spectrometry (MALDI-TOF), microscopy (SEM, Raman), and microbiological techniques (CFU, OD600, ATP-levels). With a fast and 96-well-adapted extraction protocol, mycobacteria could be inactivated and extracted for MALDI-TOF analysis. For the first time, we demonstrate growth-state-dependent changes in the mass signatures of the culture, allowing for a reliable differentiation of dormant state and exponential growth. We also demonstrate resuscitation from dormant state back to exponential growth. Viable mycobacteria were immobilized, and single organisms were analyzed individually by Raman microscopy. For single-cell Raman microscopy, Mycobacterium smegmatis cultures were fixed using a new fast and gentle single-step immobilization technique on a hydrophobic glass slide. We were able to distinguish single viable bacteria in the dormant state from their rapidly growing, genetically identical counterparts, identifying the growth state of the culture based on single-organism spectra. This allows for the separation of heterogeneous cultures depending on their growth state using the destruction-free optical method of Raman microscopy.

Pharmacokinetics, Tolerability, and Bacteriological Response of Rifampin Administered at 600, 900, and 1,200 Milligrams Daily in Patients with Pulmonary Tuberculosis.

In a multiple-dose-ranging trial, we previously evaluated higher doses of rifampin in patients for 2 weeks. The objectives of the current study were to administer higher doses of rifampin for a longer period to compare the pharmacokinetics, safety/tolerability, and bacteriological activity of such regimens. In a double-blind, randomized, placebo-controlled, phase II clinical trial, 150 Tanzanian patients with tuberculosis (TB) were randomized to receive either 600 mg (approximately 10 mg/kg of body weight), 900 mg, or 1,200 mg rifampin combined with standard doses of isoniazid, pyrazinamide, and ethambutol administered daily for 2 months. Intensive pharmacokinetic sampling occurred in 63 patients after 6 weeks of treatment, and safety/tolerability was assessed. The bacteriological response was assessed by culture conversion in liquid and solid media. Geometric mean total exposures (area under the concentration-versus-time curve up to 24 h after the dose) were 24.6, 50.8, and 76.1 mg · h/liter in the 600-mg, 900-mg, and 1,200-mg groups, respectively, reflecting a nonlinear increase in exposure with the dose (P < 0.001). Grade 3 adverse events occurred in only 2 patients in the 600-mg arm, 4 patients in the 900-mg arm, and 5 patients in the 1,200-mg arm. No significant differences in the bacteriological response were observed. Higher daily doses of rifampin (900 and 1,200 mg) resulted in a more than proportional increase in rifampin exposure in plasma and were safe and well tolerated when combined with other first-line anti-TB drugs for 2 months, but they did not result in improved bacteriological responses in patients with pulmonary TB. These findings have warranted evaluation of even higher doses of rifampin in follow-up trials. (This study has been registered at ClinicalTrials.gov under identifier NCT00760149.).

Identifying children with tuberculosis among household contacts in The Gambia.

SETTING: Greater Banjul Area of the Gambia. OBJECTIVES: To identify co-prevalent tuberculosis (TB) among child contacts of adults with smear-positive TB. DESIGN: Child contacts aged <15 years in the immediate household and compound were prospectively enrolled and evaluated for TB disease using screening questionnaires and the tuberculin skin test (TST). Symptomatic and/or TST-positive (10 mm) contacts were further investigated. RESULTS: Of 4042 child contacts who underwent symptom screening and TST, 3339 (82.6%) were diagnosed as TB-exposed but not infected, 639 (15.8%) were latently infected and 64 (1.6%) had co-prevalent TB. Of the 64 TB cases, 50 (78.1%) were from within the immediate household of the index case, and 14 (21.9%) from within the same compound. Of the 27 asymptomatic but TST-positive children diagnosed with TB, 7 were microbiologically confirmed. The median age of the TB cases was 4.4 years (interquartile range 1.9-6.9); 53.1% were aged <5 years. Of the 4042 child contacts, 206 (5%) slept in the same bed as the index case; 28.1% of all TB cases occurred in this group. Symptom screening alone would have detected only 57.8% of the co-prevalent cases. CONCLUSION: In our community setting, if contact tracing is restricted to symptom screening and immediate households only, nearly half of all co-prevalent TB disease in child contacts would be missed.

Optimising molecular diagnostic capacity for effective control of tuberculosis in high-burden settings.

The World Health Organization's 2035 vision is to reduce tuberculosis (TB) associated mortality by 95%. While low-burden, well-equipped industrialised economies can expect to see this goal achieved, it is challenging in the low- and middle-income countries that bear the highest burden of TB. Inadequate diagnosis leads to inappropriate treatment and poor clinical outcomes. The roll-out of the Xpert(®) MTB/RIF assay has demonstrated that molecular diagnostics can produce rapid diagnosis and treatment initiation. Strong molecular services are still limited to regional or national centres. The delay in implementation is due partly to resources, and partly to the suggestion that such techniques are too challenging for widespread implementation. We have successfully implemented a molecular tool for rapid monitoring of patient treatment response to anti-tuberculosis treatment in three high TB burden countries in Africa. We discuss here the challenges facing TB diagnosis and treatment monitoring, and draw from our experience in establishing molecular treatment monitoring platforms to provide practical insights into successful optimisation of molecular diagnostic capacity in resource-constrained, high TB burden settings. We recommend a holistic health system-wide approach for molecular diagnostic capacity development, addressing human resource training, institutional capacity development, streamlined procurement systems, and engagement with the public, policy makers and implementers of TB control programmes.

Isoniazid preventive treatment among child contacts of adults with smear-positive tuberculosis in The Gambia.

Setting: Greater Banjul area of The Gambia. Objectives: To evaluate uptake, adherence and completion of treatment among tuberculosis (TB) exposed children in The Gambia when isoniazid preventive treatment (IPT) is delivered at home Design: Child (age <5 years) contacts of adults with smear-positive TB were prospectively enrolled. Following symptom screening, tuberculin skin testing and clinical evaluation where indicated, those without disease were placed on daily isoniazid, provided monthly at home. Adherence was assessed by pill counts and IsoScreen™ urine test. Results: Of 404 contacts aged <5 years, 368 (91.1%) were offered IPT. Of the 328 (89.4%) for whom consent was received and who commenced IPT, 18 (5.5%) dropped out and 310 (94.5%) remained on IPT to the end of the 6-month regimen. Altogether, 255/328 children (77.7%, 95%CI 73.2-82.2) completed all 6 months, with good adherence. The IsoScreen test was positive in 85.3% (435/510) of all tests among those defined as having good adherence by pill count and in 16% (8/50) of those defined as having poor adherence (P < 0.001). A cascade of care analysis showed an overall completion rate with good adherence of 61% for all child contacts. Conclusion: Home-delivered IPT among child contacts of adults with smear-positive TB in The Gambia achieved verifiable high uptake and adherence rates. System rather than patient factors are likely to determine the success of IPT at national level.

Contexte : Région du Grand Banjul en Gambie.Objectifs : Evaluer la couverture, l'adhésion et l'achèvement du traitement parmi des enfants exposés à la tuberculose (TB) en Gambie quand le traitement préventif par isoniazide (TPI) est donné à domicile.Schéma : Les enfants âgés de <5 ans, contacts d'adultes atteints de TB à frottis positif, ont été enrôlés de manière prospective. Après dépistage sur les symptômes, test cutané à la tuberculine et évaluation clinique quand cela était indiqué, les enfants non malades ont été mis sous isoniazide, fourni une fois par mois à domicile. L'adhésion a été évaluée par un comptage des comprimés et par un test urinaire IsoScreen™.Résultats : Sur 404 contacts âgés de <5 ans, 368 (91,1%) ont été invités à bénéficier du TPI, et 328 (89,4%) ont consenti et commencé le TPI. Sur ces 328 enfants, 18 (5,5%) ont abandonné et 310 (94,5%) sont restés sous TPI jusqu'à la fin du 6e mois. Au total, 255/328 enfants (77,7% ; IC95% 73,2­82,2) ont achevé les 6 mois de traitement avec une bonne adhésion. Le test IsoScreen a été positif chez 85,3% (435/510) de tous les tests parmi ceux définis comme ayant une bonne adhésion par le comptage des comprimés et chez 16% (8/50) de ceux définis comme ayant une adhésion médiocre (P < 0,001). L'analyse de la « cascade des soins ¼ a montré, pour tous les enfants contacts, un taux de bonne adhésion d'ensemble de 61%.Conclusion: L'administration à domicile du TPI à des enfants contacts d'adultes atteints de TB à frottis positif en Gambie a abouti à une bonne couverture et à un bon taux d'adhésion, tous deux vérifiables. Ce sont les facteurs de système plutôt que ceux liés au patient qui sont susceptibles de déterminer le succès du TPI au niveau national.

Marco de referencia: La zona del Gran Banjul en Gambia.Objetivos: Evaluar la aceptación del tratamiento preventivo con isoniazida (TPI), su cumplimiento y su compleción por parte de los niños expuestos en Gambia, cuando se suministra el tratamiento en los hogares.Método: Se incluyeron en el estudio de manera prospectiva los niños menores de 5 años de edad que eran contactos de un adulto con diagnóstico de tuberculosis (TB) y baciloscopia positiva. Luego de la detección sistemática a partir de los síntomas, se practicaron la prueba cutánea de la tuberculina y la evaluación clínica cuando estaban indicadas; en caso de ausencia de enfermedad activa se inició el tratamiento diario con isoniazida, la cual se suministraba en el hogar cada mes. Se evaluó el cumplimiento en función del recuento de los comprimidos y la prueba IsoScreen™ en muestras de orina.Resultados: En los 404 contactos menores de 5 años de edad, se ofreció el TPI a 368 niños (91,1%) y 328 lo aceptaron y comenzaron a recibirlo (89,4%). De este grupo, 18 niños abandonaron el tratamiento (5,5%) y 310 recibían aun el medicamento al final del 6 mes (94,5%). De los 328 niños, 255 terminaron los 6 meses de tratamiento, con un cumplimiento satisfactorio (77,7%; IC del 95% de 73,2 hasta 82,2). La prueba IsoScreen fue positiva en el 85,3% (435/510) de los casos definidos con cumplimiento adecuado según el recuento de comprimidos y en el 16% (8/50) de los casos cuyo cumplimiento se consideró deficiente (P < 0,001). El análisis de la trayectoria asistencial reveló que en todos los contactos la tasa global de compleción con cumplimiento satisfactorio fue 61%.Conclusión: El TPI suministrado en el hogar a los niños que son contactos de un adulto con diagnóstico de TB y baciloscopia positiva alcanza altas tasas de aceptación y de cumplimiento que se pueden verificar. Los factores que determinan el éxito del TPI a escala nacional dependen del sistema de salud y no del paciente.

Ascertaining in vivo virulence of Mycobacterium tuberculosis lineages in patients in Mbeya, Tanzania.

We evaluated the relationship between the degree of immunodeficiency indicated by the number of circulating CD4+ T-cells and Mycobacterium tuberculosis lineages identified by spoligotyping and mycobacterial interspersed repetitive units-variable number of tandem repeats genotyping in human immunodeficiency virus (HIV) infected individuals with pulmonary tuberculosis from Mbeya, Tanzania. Of M. tuberculosis strains from 129 patients, respectively 55 (42.6%) and 37 (28.7%) belonged to Latin American Mediterranean and Delhi/Central-Asian lineages, while 37 (28.7%) patients were infected with other strains. There was no difference in the distribution of M. tuberculosis lineages among patients with early or advanced stages of HIV infection (P = 0.785), indicating that the virulence of strains from these lineages may not be substantially different in vivo.

Screening for pulmonary tuberculosis in a Tanzanian prison and computer-aided interpretation of chest X-rays.

SETTING: Tanzania is a high-burden country for tuberculosis (TB), and prisoners are a high-risk group that should be screened actively, as recommended by the World Health Organization. Screening algorithms, starting with chest X-rays (CXRs), can detect asymptomatic cases, but depend on experienced readers, who are scarce in the penitentiary setting. Recent studies with patients seeking health care for TB-related symptoms showed good diagnostic performance of the computer software CAD4TB. OBJECTIVE: To assess the potential of computer-assisted screening using CAD4TB in a predominantly asymptomatic prison population. DESIGN: Cross-sectional study. RESULTS: CAD4TB and seven health care professionals reading CXRs in local tuberculosis wards evaluated a set of 511 CXRs from the Ukonga prison in Dar es Salaam. Performance was compared using a radiological reference. Two readers performed significantly better than CAD4TB, three were comparable, and two performed significantly worse (area under the curve 0.75 in receiver operating characteristics analysis). On a superset of 1321 CXRs, CAD4TB successfully interpreted >99%, with a predictably short time to detection, while 160 (12.2%) reports were delayed by over 24 h with conventional CXR reading. CONCLUSION: CAD4TB reliably evaluates CXRs from a mostly asymptomatic prison population, with a diagnostic performance inferior to that of expert readers but comparable to local readers.

Contexte : La Tanzanie est lourdement frappée par la tuberculose (TB) et les prisonniers sont un groupe à haut risque qui devrait bénéficier d'un dépistage actif, comme le recommande l'Organisation Mondiale de la Santé. Les algorithmes de dépistage qui débutent par une radiographie pulmonaire peuvent détecter des cas asymptomatiques, mais ils requièrent des lecteurs de radiographies expérimentés, qui sont rares dans le contexte pénitentiaire. Des études récentes sur des patients sollicitant des soins pour des symptômes liés à la TB ont mis en évidence une bonne performance diagnostique du logiciel CAD4TB.Objectif : Evaluer le potentiel d'un dépistage assisté par ordinateur en utilisant CAD4TB au sein d'une population carcérale en majorité asymptomatique.Schéma : Étude transversale.Résultats : CAD4TB et sept professionnels de santé lisant des radiographies dans des services de TB locaux ont évalué un ensemble de 511 radiographies pulmonaires provenant de la prison d'Ukonga à Dar es Salaam et les performances ont été comparées grâce à une radiographie de référence. Deux lecteurs ont été significativement plus performants que CAD4TB, trois ont été comparables et deux ont été significativement moins bons (zone sous la courbe de 0,75 dans l'analyse ROC ­fonction d'efficacité du receveur). Sur un ensemble de 1321 radiographies pulmonaires, CAD4TB en a interprété avec succès plus de 99% avec un délai de détection prévisible court, tandis que 160 (12,2%) réponses ont été retardées de plus de 24 h avec la méthode de lecture conventionnelle.Conclusion : CAD4TB évalue de manière fiable les radiographies pulmonaires dans une population en majorité asymptomatique de détenus, avec une performance diagnostique inférieure à celle de lecteurs experts mais comparable à celle des lecteurs locaux.

Marco de referencia: Tanzania es un país con una alta tasa de morbilidad por tuberculosis (TB) y las personas en los establecimientos penitenciarios constituyen un grupo de alto riesgo de contraer la enfermedad; en esta población se debe practicar la detección sistemática activa como lo recomienda la Organización Mundial de la Salud. Los algoritmos de detección cuya etapa inicial es la radiografía de tórax pueden detectar los casos asintomáticos, pero su eficacia depende de la experiencia del profesional que interpreta las imágenes y esta competencia es escasa en los entornos penitenciarios. Algunos estudios recientes de pacientes que buscan atención sanitaria por síntomas asociados con la TB han revelado un buen rendimiento diagnóstico con la utilización del programa informático CAD4TB. Objetivo: Evaluar la utilidad de la detección sistemática de la TB asistida por el programa CAD4TB, en una población penitenciaria en su mayoría asintomática.Método: Fue este un estudio de tipo transversal.Resultados: Siete profesionales de atención sanitaria de los servicios locales de TB analizaron 511 radiografías de tórax provenientes de la prisión de Ukonga, en Dar es-Salam, con la ayuda del programa CAD4TB; se preparó un conjunto de referencia radiográfica de lectura con el fin de evaluar el rendimiento diagnóstico. El desempeño de dos de los lectores fue significativamente superior al resultado del programa CAD4TB, tres lectores obtuvieron una puntuación comparable al programa y en dos lectores se observó un rendimiento significativamente inferior (área bajo la curva: 0,75 en el análisis de eficacia diagnóstica). En un conjunto especial de 1321 radiografías de tórax el programa CAD4TB interpretó eficazmente más del 99%, con un corto lapso previsible hasta la detección, en contraste con la lectura clásica de las radiografías que dio lugar a un retraso superior a 24 horas en 160 informes (12,2%).Conclusión: El programa CAD4TB realizó una evaluación fiable de las radiografías provenientes de una población penitenciaria en su mayor parte asintomática. El rendimiento diagnóstico del programa fue inferior al rendimiento de los lectores expertos, pero comparable con el rendimiento de los lectores locales.

A continuously monitored colorimetric method for detection of Mycobacterium tuberculosis complex in sputum.

SETTING: Mbeya, Tanzania. OBJECTIVE: To develop a new liquid culture method to detect Mycobacterium tuberculosis complex (MTC) in sputum using 2,3-diphenyl-5-thienyl-(2)-tetrazolium (STC), the nitrate reductase assay (NRA) and p-nitrobenzoic acid (PNB). DESIGN: Ninety-three sputum samples collected from 18 tuberculosis patients were decontaminated with N-acetyl-L-cysteine-sodium hydroxide using MGIT™ 960 and in STC-NRA cultures, both in the presence and in the absence of PNB, an inhibitor of MTC growth. The reduction of STC by colour change indicated mycobacterial growth; NRA was then performed to confirm MTC. RESULTS: STC-NRA culture was positive for acid-fast bacilli in 66/93 (71%) samples, of which 60/93 (64.5%) were identified as MTC-positive and 6/93 (6.5%) as indeterminate mycobacteria. MGIT indicated MTC in 59/93 (63.4%) cultures. Contamination was detected in 12/93 (13%) STC-NRA cultures vs. 29/93 (31.2%) MGIT cultures. The mean time to detection (TTD) of MTC using STC-NRA was 14 days and 7 days using MGIT. CONCLUSION: The STC-NRA method is sensitive for the detection of MTC in sputum. TTD increased with duration of anti-tuberculosis treatment, highlighting the value of this method in monitoring treatment success. The method is simple and inexpensive and, unlike MGIT, does not require technical equipment. The preliminary performance characteristics of the method should be further evaluated in larger studies.

Performance of the Xpert® MTB/RIF assay in an active case-finding strategy: a pilot study from Tanzania.

In this pilot study, we evaluated the Xpert® MTB/RIF assay in an active case-finding strategy, using two spot sputum samples collected within a 1-hour interval from household contacts of smear-positive TB index cases. Tuberculosis (TB) confirmed by culture served as the reference standard. Among 219 enrolled contacts, the yield of active TB was 2.3%. While the sensitivity of smear microscopy was 60% (95%CI 14.7-94.7), Xpert MTB/RIF achieved a sensitivity of 100% (95%CI 47.81-100.0). All culture-confirmed cases tested positive by Xpert MTB/RIF on the first submitted sample, suggesting that the evaluation of only one sample could be sufficient for TB diagnosis in this context.

Detecting 2009 pandemic influenza A (H1N1) virus infection: availability of diagnostic testing led to rapid pandemic response.

Diagnostic tests for detecting emerging influenza virus strains with pandemic potential are critical for directing global influenza prevention and control activities. In 2008, the Centers for Disease Control and Prevention received US Food and Drug Administration approval for a highly sensitive influenza polymerase chain reaction (PCR) assay. Devices were deployed to public health laboratories in the United States and globally. Within 2 weeks of the first recognition of 2009 pandemic influenza H1N1, the Centers for Disease Control and Prevention developed and began distributing a new approved pandemic influenza H1N1 PCR assay, which used the previously deployed device platform to meet a >8-fold increase in specimen submissions. Rapid antigen tests were widely used by clinicians at the point of care; however, test sensitivity was low (40%-69%). Many clinical laboratories developed their own pandemic influenza H1N1 PCR assays to meet clinician demand. Future planning efforts should identify ways to improve availability of reliable testing to manage patient care and approaches for optimal use of molecular testing for detecting and controlling emerging influenza virus strains.

Evaluation of Diagnos TB AG, a flow-through immunoassay for rapid detection of pulmonary tuberculosis.

We evaluated the diagnostic performance of the Diagnos TB AG immunoassay in 171 Tanzanians with suspected pulmonary tuberculosis (TB). The sensitivity and specificity, and positive and negative predictive values of the rapid test for the detection of pulmonary TB in this population were respectively 60.0%, 33.3%, 40.3% and 52.6%. In its current configuration, this test will not help overcome difficulties in the rapid diagnosis of TB.

New pre-pandemic influenza vaccines: an egg- and adjuvant-independent human adenoviral vector strategy induces long-lasting protective immune responses in mice.

Highly pathogenic avian H5N1 influenza viruses that are currently circulating in southeast Asia may acquire the potential to cause the next influenza pandemic. A number of alternate approaches are being pursued to generate cross-protective, dose-sparing, safe, and effective vaccines, as traditional vaccine approaches, i.e., embryonated egg-grown, are not immunogenic. We developed a replication-incompetent adenoviral vector-based, adjuvant- and egg-independent pandemic influenza vaccine strategy as a potential alternative to conventional egg-derived vaccines. In this paper, we address suboptimal dose and longevity of vaccine-induced protective immunity and demonstrate that a vaccine dose as little as 1 x 10(6) plaque-forming unit (PFU) is sufficient to induce protective immune responses against a highly pathogenic H5N1 virus. Furthermore, the vaccine-induced humoral and cellular immune responses and protective immunity persisted at least for a year.

In-depth analysis of a heterosexually acquired human immunodeficiency virus type 1 superinfection: evolution, temporal fluctuation, and intercompartment dynamics from the seronegative window period through 30 months postinfection.

Human immunodeficiency virus type 1 (HIV-1) superinfection refers to the acquisition of another strain by an already infected individual. Here we report a comprehensive genetic analysis of an HIV-1 superinfection acquired heterosexually. The infected individual was in a high-risk cohort in Tanzania, was exposed to multiple subtypes, and was systematically evaluated every 3 months with a fluorescent multi-region genotyping assay. The subject was identified in the window period and was first infected with a complex ACD recombinant strain, became superinfected 6 to 9 months later with an AC recombinant, and was monitored for >2.5 years. The plasma viral load exceeded 400,000 copies/ml during the first 9 months of infection but resolved to the set point of 67,000 copies/ml by 3 months after superinfection; the CD4 cell count was 377 cells/mul at 30 months. Viral diversity was evaluated with techniques designed to fully sample the quasi-species, permitting direct observation of the evolution, temporal fluctuation, and intercompartment dynamics of the initial and superinfecting strains and recombinants derived from them. Within 3 months of superinfection, seven different molecular forms were detected in gag and six were detected in env. The proportions of forms fluctuated widely over time in plasma and peripheral blood mononuclear cells, illustrating how challenging the detection of dually infected individuals can be. Strain-specific nested PCR confirmed that the superinfecting strain was not present until the 9 month follow-up. This study further defines the parameters and dynamics of superinfection and will foster appropriate studies and approaches to gain a more complete understanding of risk factors for superinfection and its impact on clinical progression, epidemiology, and vaccine design.