Serum and cerebrospinal fluid host proteins indicate stroke in children with tuberculous meningitis

Introduction Stroke is a common complication in children with tuberculous meningitis (TBM). Host proteins may give us insight into the mechanisms of stroke in TBM and serve as biomarkers for detection of stroke, however, they have not been widely explored. In this study, we compared the concentrations of cerebrospinal fluid (CSF) and serum proteins between children who had TBM-related stroke and children with TBM without stroke. Methods We collected CSF and serum from 47 children consecutively admitted to the Tygerberg Academic Hospital in Cape Town, South Africa between November 2016, and November 2017, on suspicion of having TBM. A multiplex platform was used to measure the concentrations of 69 host proteins in CSF and serum from all study participants. Results After classification of study participants, 23 (48.9%) out of the 47 study participants were diagnosed with TBM, of which 14 (60.9%) demonstrated radiological arterial ischemic infarction. The levels of lipocalin-2, sRAGE, IP-10/ CXCL10, sVCAM-1, MMP-1, and PDGF-AA in CSF samples and the levels of D-dimer, ADAMTS13, SAA, ferritin, MCP-1/ CCL2, GDF-15 and IL-13 in serum samples were statistically different between children who had TBM-related stroke and children with TBM without stroke. After correcting for multiple testing, only the levels of sVCAM-1, MMP-1, sRAGE, and IP-10/ CXCL10 in CSF were statistically different between the two groups. CSF and serum protein biosignatures indicated stroke in children diagnosed with TBM with up to 100% sensitivity and 88.9% specificity. Conclusion Serum and CSF proteins may serve as biomarkers for identifying individuals with stroke amongst children diagnosed with TBM at admission and may guide us to understand the biology of stroke in TBM. This was a pilot study, and thus further investigations in larger studies are needed

Prospective evaluation of host biomarkers other than interferon gamma in QuantiFERON Plus supernatants as candidates for the diagnosis of tuberculosis in symptomatic individuals

CITATION: Manngo, P. M. et al. 2019. Prospective evaluation of host biomarkers other than interferon gamma in QuantiFERON Plus supernatants as candidates for the diagnosis of tuberculosis in symptomatic individuals. Journal of Infection, 79(3):228-235, doi:10.1016/j.jinf.2019.07.007.The original publication is available at https://www.journalofinfection.comBackground: There is an urgent need for new tools for the diagnosis of TB. We evaluated the usefulness recently described host biomarkers in supernatants from the newest generation of the QuantiFERON test (QuantiFERON Plus) as tools for the diagnosis of active TB. Methods: We recruited individuals presenting at primary health care clinics in Cape Town, South Africa with symptoms requiring investigation for TB disease, prior to the establishment of a clinical diagnosis. Participants were later classified as TB or other respiratory diseases (ORD) based on the results of clinical and laboratory tests. Using a multiplex platform, we evaluated the concentrations of 37 host biomarkers in QuantiFERON Plus supernatants from study participants as tools for the diagnosis of TB. Results: Out of 120 study participants, 35(29.2%) were diagnosed with active TB, 69(57.5%) with ORD whereas 16(13.3%) were excluded. 14(11.6%) of the study participants were HIV infected. Although individ- ual host markers showed potential as diagnostic candidates, the main finding of the study was the identi- fication of a six-marker biosignature in unstimulated supernatants (Apo-ACIII, CXCL1, CXCL9, CCL8, CCL-1, CD56) which diagnosed TB with sensitivity and specificity of 73.9%(95% CI; 51.6–87.8) and 87.6%(95% CI; 77.2–94.5), respectively, after leave-one-out cross validation. Combinations between TB-antigen specific biomarkers also showed potential (sensitivity of 77.3% and specificity of 69.2%, respectively), with multi- ple biomarkers being significantly different between TB patients, Quantiferon Plus Positive and Quantif- eron Plus negative individuals with ORD, regardless of HIV status. Conclusions: Biomarkers detected in QuantiFERON Plus supernatants may contribute to adjunctive diag- nosis of TB.EDCTP , grant no: DRIA2014-311National Research FoundationICIDR (grant no: 5U01IA115619)Publisher's versio

Evaluation of the potential of Mycobacterium tuberculosis antigen-specific host biomarkers detected in QuantiFERON® TB GOLD Plus supernatants in the diagnosis of TB disease

Thesis (MSc)--Stellenbosch University, 2018.ENGLISH ABSTRACT: BACKGROUND The diagnosis of tuberculosis (TB) disease remains a challenge. This is mainly due to limitations with the current TB diagnostic tests including unavailability of rapid pointof-care tests. New TB diagnostic tests are therefore urgently needed. The QuantiFERON-TB® Gold (QFT) Plus test is a recently introduced test for the diagnosis of M. tb infection, and disease in some patient groups. As this is a relatively new test which is currently in use worldwide, it is important that its performance be evaluated, especially in high TB burden settings. Furthermore, it is not known whether measurement of host markers other than Interferon-gamma in culture supernatants of individuals with active TB or other respiratory diseases (ORD), has potential in the diagnosis of TB disease. OBJECTIVES 1) To evaluate the usefulness of the QFT Plus test in the diagnosis of TB disease, and assess the utility of the test, when used in combination with symptoms, as a tool for diagnosis of TB disease in people suspected of having active TB in a high burden setting. 2) To evaluate alternative host biomarkers detected in QFT Plus supernatants, other than IFN-γ as biosignatures for the diagnosis of active TB METHODS We recruited 120 participants presenting at a primary health care clinics in Cape Town, South Africa with symptoms requiring investigation for TB disease. These participants formed part of a larger ongoing biomarker project known as the ‘ScreenTB’ study. Participants were later classified as TB or ORD based on the results of clinical and laboratory tests. After performing the standard QFT Plus test in study participants, the concentrations of 37 host biomarkers were evaluated in culture supernatants using a multiplex immunoassay. RESULTS Out of 120 individuals included in the study, 35 (29.2%) were diagnosed with active TB and were culture positive. The QFT Plus test diagnosed TB disease in all study participants with sensitivity and specificity >70%. A combination of symptoms including cough, fever and weight loss diagnosed TB disease with sensitivity and specificity >70% with an area under the receiver operator characteristics curve of 0.81. Multiple host biomarkers detected in the unstimulated and antigen-stimulated QFT Plus tubes showed potential as diagnostic markers for TB. Individual markers which diagnosed TB disease with sensitivities and specificities >60% included ITAC-1, IL-3, I-309, MIG, and EGF, P-selectin. Combinations between host biomarkers showed potential in the diagnosis of TB disease with a six-marker biosignature derived from unstimulated supernatants (APO-CII, ITAC-1, MIG, MCP-2, I-309, and NCAM-1) diagnosing TB disease with a sensitivity and specificity >78%, a four-marker TB1 and TB2 antigenspecific biosignature (TNFα, LIGHT, MIG and P-selectin ) which diagnosed TB disease with sensitivity and specificity >73%, after leave-one-out cross validation. CONCLUSION The sensitivity of the QFT Plus test for active TB was inferior to the published >80% mentioned in the package insert by the manufacturer. Host biomarkers detected in QFT Plus supernatants showed potential in the diagnosis of active TB disease. Further validation studies are needed before such markers may be considered as candidate biomarkers for a blood-based diagnostic test for active TB.AFRIKAANSE OPSOMMING: AGTERGROND Die diagnose van tuberkulose (TB siekte) bly 'n uitdaging. Dit is hoofsaaklik te wyte aan beperkinge met die huidige TB diagnostiese toetse, insluitend die beskikbaarheid van vinnige punt van soegtoetse. Nuwe TB diagnostiese toetse is dus dringend nodig. Die QuantiFERON-TB® Goud (QFT) Plus toets is 'n onlangs ontwikkelde toets vir die diagnose van M. tb infeksie en tuberkulose in sommige pasiëntgroepe. Aangesien dit 'n relatief nuwe toets is wat tans wêreldwyd gebruik word, is dit belangrik dat die prestasie vd toets geëvalueer word, veral in hoë TB-lasinsweld dtrelie tellings. Verder is dit nie bekend of meting van gasheermerkers behalwe Interferon-gamma in kweek supernatante van individue met aktiewe TB of ander respiratoriese siektes (ORD), potensiaal het in die diagnose van TB-siekte. DOELWITTE 1) Om die nut van die QFT Plus-toets in die diagnose van TB-siekte te evalueer, wanneer dit in kombinasie met simptome gebruik word, as 'n instrument vir die diagnose van TB-siekte by mense wat vermoed word dat hulle aktiewe TB heit in 'n hoë lastrehe. 2) Om alternatiewe gasheerbiomerkers wat in QFT Plus supernatante aangetref word, te evalueer, anders as IFN-γ as biomeker vir die diagnose van aktiewe TB METODES Ons het 120 deelnemers gewerf by 'n primêre gesondheidsorgkliniek in Kaapstad, Suid-Afrika, met simptome wat ondersoek na TB-siekte vereis. Hierdie deelnemers het deel gevorm van 'n groter voortgesette biomerkerprojek wat bekend staan as die 'ScreenTB'-studie. Deelnemers is geklassifiseer as TB of ORD gebaseer op die resultate van kliniese en laboratoriumtoetse. Nadat die standaard QFT Plus-toets in studie-deelnemers uitgevoer is, is die konsentrasies van 37 gasheerbiomerkers geëvalueer in kweek supernatante met behulp van 'n veelvuldige imuuufoetse. RESULTATE Uit 120 individue wat in die studie ingesluit is, is 35 (29,2%) met aktiewe TB gediagnoseer en was kultuur positief. Die QFT Plus-toets het TB-siektes in alle studiedeelnemers met 'n sensitiwiteit en spesifisiteit van> 70% gediagnoseer. 'n Kombinasie van simptome soos hoes, koors en gewigsverlies diagnoseer TB siekte met sensitiwiteit en spesifisiteit> 70% met 'n gebied onder die ontvanger operateur eienskappe kurwe van 0.81. Veelvuldige gasheerbiomerkers wat in die ongestimuleerde en antigeen-gestimuleerde QFT Plus-buise opgespoor is, het potensiaal as diagnostiese merkers vir TB vertoon. Individuele merkers wat TB-siekte gediagnoseer het met sensitiwiteit en spesifieke eienskappe> 60% sluit in ITAC-1, IL3, I-309, MIG en EGF, P-selektien. Kombinasies tussen gasheerbiomarkers het potensiaal getoon in die diagnose van TB-siekte. Ses biomakers van ongestimuleerde supernatante (APO-CII, ITAC-1, MIG, MCP-2, I-309, en NCAM-1) het 'n sensitiwiteit en spesifisiteit getoon van >78%. Die vier-biomeker TB1 en TB2 antigenspesifieke kombinasie (TNFa, LIG, MIG en P-selektien) het TB slette gecliagnoseer met 'n sensitiwiteit en spesifisileit >73% na verlof-een-uit kruis validasie. AFSLUITING Die sensitiwiteit van die QFT Plus-toets vir aktiewe TB was nie soos die gepubliseerde> 80% wat in die pakketstuk deur die vervaardiger genoem word nie. Gasheer biomerkers wat in QFT Plus supernatante aangetoon is, het potensiaal getoon in die diagnose van aktiewe TB-siekte. Verdere valideringstudies is nodig voordat sulke merkers as kandidaat-biomerkers beskou kan word vir 'n bloedgebaseerde diagnostiese toets vir aktiewe TB.Master

Dexmedetomidine premedication for fiberoptic intubation in patients of temporomandibular joint ankylosis: A randomized clinical trial

Background : Fiberoptic intubation is the gold standard technique for difficult airway management in patients of temporomandibular joint. This study was aimed to evaluate the clinical efficacy and safety of dexmedetomidine as premedication with propofol infusion for fiberoptic intubation. Methods: Consent was obtained from 46 adult patients of temporomandibular joint ankylosis, scheduled for gap arthroplasty. They were enrolled for thisdouble-blind, randomized, prospective clinical trial with two treatment groups - Group D and Group P, of 23 patients each. Group D patients had received premedication of dexmedetomidine 1 μg/kg infused over 10 min followed by sedative propofol infusion and the control Group P patients were given only propofol infusion to achieve sedation. Condition achieved at endoscopy, intubating conditions, hemodynamic changes and postoperative events were evaluated as primary outcome. Results : The fiberoptic intubation was successful with satisfactory endoscopic and intubating condition in all patients. Dexmedetomidine premedication has provided satisfactory conditions for fiberoptic intubation and attenuated the hemodynamic response of fiberoptic intubation than the propofol group. Conclusion : Fiberoptic intubation was found to be easier with dexmedetomidine premedication along with sedative infusion of propofol with complete amnesia of the procedure, hemodynamic stability and preservation of patent airway

Potential of host serum protein biomarkers in the diagnosis of tuberculous meningitis in children

CITATION: Manyelo, C. M., et al. 2019. Potential of host serum protein biomarkers in the diagnosis of tuberculous meningitis in children. Frontiers in Pediatrics, 7:376, doi:10.3389/fped.2019.00376.The original publication is available at https://www.frontiersin.orgPublication of this article was funded by the Stellenbosch University Open Access Fund.Background: Tuberculous meningitis (TBM) is the most severe form of tuberculosis and results in high morbidity and mortality in children. Diagnostic delay contributes to the poor outcome. There is an urgent need for new tools for the rapid diagnosis of TBM, especially in children. Methods: We collected serum samples from children in whom TBM was suspected at a tertiary hospital in Cape Town, South Africa. Children were subsequently classified as having TBM or no TBM using a published uniform research case-definition. Using a multiplex cytokine array platform, we investigated the concentrations of serum biomarkers comprising biomarkers that were previously found to be of value in the diagnosis of adult pulmonary TB (CRP, SAA, CFH, IFN-γ, IP-10, Apo-AI, and transthyretin) plus other potentially useful host biomarkers as diagnostic candidates for TBM. Findings: Out of 47 children included in the study, 23 (48.9%) had a final diagnosis of TBM and six were HIV infected. A modified version of the adult 7-marker biosignature in which transthyretin was replaced by NCAM1, diagnosed TBM in children with AUC of 0.80 (95% CI, 0.67–0.92), sensitivity of 73.9% (95% CI, 51.6–89.8%) and specificity of 66.7% (95% CI, 44.7–84.4%), with the other six proteins in the signature (CRP, IFN-γ, IP-10, CFH, Apo-A1, and SAA) only achieving and AUC of 0.75 (95% CI, 0.61–0.90) when used in combination. A new childhood TBM specific 3-marker biosignature (adipsin, Aβ42, and IL-10) showed potential in the diagnosis of TBM, with AUC of 0.84 (95% CI, 0.73–0.96), sensitivity of 82.6% (95 CI, 61.2–95.0%) and specificity of 75.0% (95% CI, 53.3–90.2%) after leave-one-out cross validation. Conclusion: A previously described adult 7-marker serum protein biosignature showed potential in the diagnosis of TBM in children. However, a smaller childhood TBM-specific 3-marker signature demonstrated improved performance characteristics. Our data indicates that blood-based biomarkers may be useful in the diagnosis of childhood TBM and requires further validation in larger cohort studies.https://www.frontiersin.org/articles/10.3389/fped.2019.00376/fullPublisher's versionAuthors retain copyrigh

Application of cerebrospinal fluid host protein biosignatures in the diagnosis of tuberculous meningitis in children from a high burden setting

CITATION: Manyelo, C. M., et al. 2019. Application of cerebrospinal fluid host protein biosignatures in the diagnosis of tuberculous meningitis in children from a high burden setting. Mediators of Inflammation, 2019 (Article ID 7582948), doi:10.1155/2019/7582948.The original publication is available at https://www.hindawi.comPublication of this article was funded by the Stellenbosch University Open Access FundBackground. The diagnosis of tuberculous meningitis (TBM) especially in children is challenging. New tests are urgently needed for the diagnosis of the disease, especially in resource-limited settings. Methods. We collected cerebrospinal fluid (CSF) samples from children presenting with symptoms requiring investigation for meningitis at a tertiary hospital in Cape Town, South Africa. Children were later classified as TBM or no TBM using published case definitions. Using a multiplex platform, we investigated the concentrations of biomarkers comprising a previously established 3-marker biosignature (VEGF, IL-13, and LL-37) and other potentially useful host biomarkers as diagnostic candidates for TBM. Findings. Out of 47 children, age, 3 months to 13 years, 23 were diagnosed with TBM and six (16%) were HIV-infected. We validated the previously identified CSF biosignature (sensitivity of 95.7% (95% CI, 79.0-99.2%) and specificity of 37.5% (95% CI, 21.2-57.3%)). However, substitution of IL-13 and LL-37 with IFN-γ and MPO, respectively, resulted in improved accuracy (area under the ROC curve (AUC) = 0 97, 95% CI, 0.92-1.00, up to 91.3% (21/23) sensitivity and up to 100% (24/24) specificity). An alternative four-marker biosignature (sICAM-1, MPO, CXCL8, and IFN-γ) also showed potential, with an AUC of 0.97. Conclusion. We validated a previously identified CSF biosignature and showed that refinement of this biosignature by incorporation of other biomarkers diagnosed TBM with high accuracy. Incorporation of these biomarkers into a point-of-care or bedside diagnostic test platform may result in the improved management of TBM in children.https://www.hindawi.com/journals/mi/2019/7582948/Publisher's versio