Screening and identification of serum biomarkers of osteoarticular tuberculosis based on mass spectrometry.

BACKGROUND: In view of the current difficulty of clinically diagnosing osteoarticular tuberculosis, our aim was to use mass spectrometry to establish diagnostic models and to screen and identify serum proteins which could serve as potential diagnostic biomarkers for early detection of osteoarticular tuberculosis. METHODS: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to select an osteoarticular tuberculosis-specific serum peptide profile and establish diagnostic models. Further, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to identify potential serum biomarkers that could be used for auxiliary diagnosis of osteoarticular tuberculosis, and then clinical serum samples were used to verify these biomarkers by enzyme-linked immunosorbent assay (ELISA). RESULTS: We established four diagnostic models that can distinguish osteoarticular tuberculosis from rheumatoid arthritis, ankylosing spondylitis, osteoarticular infections, and healthy adults. The models were osteoarticular tuberculosis-rheumatoid arthritis, osteoarticular tuberculosis-ankylosing spondylitis, osteoarticular tuberculosis-osteoarticular infections, and osteoarticular tuberculosis-healthy adult, and their accuracy was 76.78%, 79.02%, 83.77%, and 88.16%, respectively. Next, we selected and identified 18 proteins, including complement factor H-related protein 1 (CFHR1) and complement factor H-related protein 2 (CFHR2), which were upregulated in the tuberculosis group only. CONCLUSIONS: We successfully established four diagnostic models involving osteoarticular tuberculosis, rheumatoid arthritis, ankylosing spondylitis, osteoarticular infections, and healthy adults. Furthermore, we found that CFHR1 and CFHR2 may be two valuable auxiliary diagnostic indicators for osteoarticular tuberculosis. These results provide reference values for rapid and accurate diagnosis of osteoarticular tuberculosis.

Diagnostic value of ELISPOT technique for osteoarticular tuberculosis.

BACKGROUND: Osteoarticular tuberculosis (TB) is a common severe form of extrapulmonary tuberculosis. Early di- agnosis and treatment can decrease the deformity of spine and limbs and joint dysfunction. METHODS: We compared and evaluated two commercially available rapid test kits based on the ELISPOT assay for the diagnosis of osteoarticular disease. RESULTS: The diagnostic sensitivity and specificity of the FS-SPOT assay (50.0% and 85.7%) were similar to those of the T-SPOT-TB assay (45.5% and 81.0%). When the two test wells in the T-SPOT-TB assay were both positive, the test wells in FS-SPOT assay were usually positive with the number of SFCs exceeding those in the negative control wells by more than 30. The sensitivities, specificities, PPV, NPV, and agreement of FS-SPOT assay results in 99 TB cases and 54 non-TB disease cases were 55.6%, 83.3%, 84.7%, 52.9%, and 66.0%, respectively. SFC counts from test wells in the TB group were significantly higher than those from the non-TB group (p < 0.001). CONCLUSIONS: Higher numbers of SFCs in the ELISPOT assay suggest higher risk of active TB. ELISPOT may be a diagnostic aide for active osteoarticular TB.

Serum Protein Electrophoresis Patterns in Patients with Osteoarticular Tuberculosis.

CONTEXT AND AIMS: Tuberculosis (TB) is a leading cause of infectious disease deaths in India. It is also one of the most challenging diseases to diagnose and treat effectively. TB can occur both in the lungs and in extrapulmonary locations through hematogenous spread. Osteoarticular TB is a type of extrapulmonary characterized by atypical presentation. If diagnosed early, it can be treated effectively with reduced risk of mortality. SUBJECTS AND METHODS: At Nalanda medical college and hospital, an 18-month prospective research was undertaken. The study included a total of 120 patients with osteoarticular TB. Serum electrophoresis of blood samples was performed at baseline, 2 months, and 4 months following antitubercular medication administration. The fractions of albumin, α1, α2, ß, and γ globulins were estimated and compared with the baseline value. RESULTS: It was observed that as the disease progressed and became more chronic, there was a decrease in albumin and an increase in α1, α2, ß, and γ globulin percentages of serum proteins. Upon follow-up, the serum electrophoresis revealed that these values observed during baseline could be reversed by the administration of antitubercular drugs. CONCLUSIONS: This study suggests that analyzing serum protein fractions could be a cost-effective strategy to determine the presence of osteoarticular TB and also aid in initiating antitubercular treatment.

RésuméContexte et objectifs: La tuberculose (TB) est l'une des principales causes de décès par maladies infectieuses en Inde. C'est également l'une des maladies les plus difficiles à diagnostiquer et à traiter efficacement. La tuberculose peut survenir à la fois dans les poumons et dans des localisations extrapulmonaires par propagation hématogène. La tuberculose ostéoarticulaire est un type de tuberculose extrapulmonaire caractérisée par une présentation atypique. Si elle est diagnostiquée tôt, elle peut être traitée efficacement avec un risque de mortalité réduit. Sujets et méthodes: À la faculté de médecine et à l'hôpital de Nalanda, une recherche prospective de 18 mois a été entreprise. L'étude a inclus un total de 120 patients atteints de tuberculose ostéoarticulaire. L'électrophorèse sérique des échantillons de sang a été réalisée au départ, 2 mois et 4 mois après l'administration de médicaments antituberculeux. Les fractions d'albumine, a1, a2, b et g globulines ont été estimées et comparées à la valeur de base. Résultats: Il a été observé qu'à mesure que la maladie progressait et devenait plus chronique, il y avait une diminution de l'albumine et une augmentation des pourcentages de globulines a1, a2, b et g des protéines sériques. Lors du suivi, l'électrophorèse sérique a révélé que ces valeurs observées au départ pouvaient être inversées par l'administration de médicaments antituberculeux. Conclusions : Cette étude suggère que l'analyse des fractions protéiques sériques pourrait constituer une stratégie rentable pour déterminer la présence d'une tuberculose ostéoarticulaire et également faciliter l'initiation d'un traitement antituberculeux.

[Evaluation using QFT-2G/TBGL/LAM in the patients with osteoarticular tuberculosis of post-treatment period].

PURPOSE: To evaluate results of QFT-2G/TBGL/ LAM in patients who had been completed the antituberculosis treatment for osteoarticular tuberculosis with various periods after the completion of the treatment, MATERIALS AND METHODS: Fifty-five patients who had been completed the antituberculosis treatment for osteoarticular tuberculosis at least one year after the completion of treatment were evaluated using QFT-2G/TBGL/LAM tests. Forty patients with spinal tuberculosis and 15 patients with articular tuberculosis were included. The patients with the period after the completion of the treatment less than 4 years were classifled as short-term group (33 patients) and those with the period not less than 4 years were classified as mid-long-term group (22 patients). The results of the tests were compared between the two groups. RESULTS: The result of QFT-2G test was positive in 60.6% of the patients in short-term group while 45.5% in mid-long-term group (p=0.12). On the other hand, the result of TBGL test was positive in 75.8% of the patients in short-term group whereas 22.7% in mid-long-term group (p=0.0001) and the result of LAM test was positive in 90.9% of the patients in short-term group whereas 63.6% in mid-long-term group (p= 0.01), both of these tests showed significantly low,er positive rate in mid-long-term group. There was no significant difference in the comparisons between patient groups writh/without pulmonary tuberculosis as well as with/without surgical treatment. CONCLUSION: The patients with a history of osteoarticular tuberculosis tend to show positive results of QFT-2-G test for a prolonged period, whereas significantly less positive results of TBGL/LAM tests in mid-long-term.

Diagnostic values of peripheral blood T-cell spot of tuberculosis assay (T-SPOT.TB) and magnetic resonance imaging for osteoarticular tuberculosis: a case-control study.

OBJECTIVE: Early diagnosis of osteoarticular tuberculosis helps improve patients' outcomes, but little is known about the accuracy of noninvasive diagnostic methods. This case-control study aimed to assess the diagnostic value of peripheral blood T-cell spot of tuberculosis assay (T-SPOT.TB) and magnetic resonance imaging (MRI). METHODS: Patients suspected with osteoarticular tuberculosis were retrospectively included and diagnosed according to the composite reference standard. T-SPOT.TB was used to detect the number of cells secreting Interferon gamma. Diagnostic performance of T-SPOT.TB and MRI alone and combined were evaluated. RESULTS: Among the suspected patients, 92 had osteoarticular tuberculosis and 137 non- osteoarticular tuberculosis. T-SPOT.TB assay alone had a higher sensitivity (0.73 vs. 0.60) but a lower specificity (0.69 vs. 0.91 P>0.05) in diagnosing osteoarticular tuberculosis. Combined serial test showed a sensitivity and specificity 0.47, 0.97, respectively, whereas combined parallel test showed a sensitivity and specificity of 0.86, 0.65, respectively. Specificity was higher in the combined serial test than in the T-SPOT.TB assay (P=0.007) or MRI alone (P < 0.001). Furthermore, sensitivity was higher in the combined parallel test than in the T-SPOT.TB assay (P < 0.001) or MRI alone (P < 0.001). CONCLUSIONS: Combined blood T-cell spot of tuberculosis assay and osteoarticular MRI have higher sensitivity and specificity for noninvasive osteoarticular tuberculosis diagnosis, compared with either method alone.

Immune status changing helps diagnose osteoarticular tuberculosis.

OBJECTIVE: This study is aimed to develop a new nomogram for the clinical diagnosis of osteoarticular tuberculosis (TB). METHODS: xCell score estimation to obtained the immune cell type abundance scores. We downloaded the expression profile of GSE83456 from GEO and proceed xCell score estimation. The routine blood examinations of 326 patients were collected for further validation. We analyzed univariate and multivariate logistic regression to identified independent predicted factor for developing the nomogram. The performance of the nomogram was assessed using the receiver operating characteristic (ROC) curves. The correlation of ESR with lymphocytes, monocytes, and ML ratio was performed and visualized in osteoarticular TB patients. RESULTS: Compared with the healthy control group in the dataset GSE83456, the xCell score of basophils, monocytes, neutrophils, and platelets was higher, while lymphoid was lower in the EPTB group. The clinical data showed that the cell count of monocytes were much higher, while the cell counts of lymphocytes were lower in the osteoarticular TB group. AUCs of the nomogram was 0.798 for the dataset GSE83456, and 0.737 for the clinical data. We identified the ML ratio, BMI, and ESR as the independent predictive factors for osteoarticular TB diagnosis and constructed a nomogram for the clinical diagnosis of osteoarticular TB. AUCs of this nomogram was 0.843. CONCLUSIONS: We demonstrated a significant change between the ML ratio of the EPTB and non-TB patients. Moreover, we constructed a nomogram for the clinical diagnosis of the osteoarticular TB diagnosis, which works satisfactorily.

Tuberculosis infection in patients with systemic lupus erythematosus: pulmonary and extra-pulmonary infection compared.

Systemic lupus erythematosus (SLE) patients had an increased susceptibility to tuberculosis (TB). The aim of this study was to investigate the prevalence and clinical characteristics of TB in SLE patients, with focus on the differences between pulmonary and extra-pulmonary TB. This is a retrospective study that reviewed the medical records of 3,179 SLE patients from 1985 to 2004. The diagnosis of TB was confirmed by one of the following: positive acid-fast bacillus (AFB) smear, positive culture of Mycobacterium tuberculosis from appropriate specimens, or a histopathologic finding of caseating granuloma on specimen. During the 20-year review period, TB was documented in 19 SLE patients, with 21 episodes. Ten of 21 episodes (47.6%) were pulmonary TB while the other 11 episodes (52.4%) were extra-pulmonary TB. Among extra-pulmonary TB, there were joint and cutaneous involvements in five, miliary in two, Pott's disease in two, peritoneum in one, and spleen in one. The most common manifestations of TB were fever and cough. Delayed diagnosis and adverse effects of anti-TB therapy were observed in the extra-pulmonary TB group. While SLE patients commonly present with prolonged fever or chronic cough, tuberculosis infection should be taken into consideration.

The use of immunomodulators as an adjunct to antituberculous chemotherapy in non-responsive patients with osteo-articular tuberculosis.

We studied 51 patients with osteo-articular tuberculosis who were divided into two groups. Group I comprised 31 newly-diagnosed patients who were given first-line antituberculous treatment consisting of isoniazid, rifampicin, ethambutol and pyrazinamide. Group II (non-responders) consisted of 20 patients with a history of clinical non-responsiveness to supervised uninterrupted antituberculous treatment for a minimum of three months or a recurrence of a previous lesion which on clinical observation had healed. No patient in either group was HIV-positive. Group II were treated with an immunomodulation regime of intradermal BCG, oral levamisole and intramuscular diphtheria and tetanus vaccines as an adjunct for eight weeks in addition to antituberculous treatment. We gave antituberculous treatment for a total of 12 to 18 months in both groups and they were followed up for a mean of 30.2 months (24 to 49). A series of 20 healthy blood donors served as a control group.Twenty-nine (93.6%) of the 31 patients in group I and 14 of the 20 (70%) in group II had a clinicoradiological healing response to treatment by five months. The CD4 cell count in both groups was depressed at the time of enrolment, with a greater degree of depression in the group-II patients (686 cells/mm(3) (sd 261) and 545 cells/mm(3) (sd 137), respectively; p < 0.05). After treatment for three months both groups showed significant elevation of the CD4 cell count, reaching a level comparable with the control group. However, the mean CD4 cell count of group II (945 cells/mm(3) (sd 343)) still remained lower than that of group I (1071 cells/mm(3) (sd 290)), but the difference was not significant. Our study has shown encouraging results after immunomodulation and antituberculous treatment in non-responsive patients. The pattern of change in the CD4 cell count in response to treatment may be a reliable clinical indicator.

Lymphocyte/monocyte ratio in osteoarticular tuberculosis in children: a haematological biomarker revisited.

INTRODUCTION: *Work attributed to Department of Pediatric Orthopaedics, Chacha Nehru Bal Chikitsalaya, Geeta Colony, Delhi-110031(India)This study tested the hypothesis whether the lymphocyte/monocyte ratio (L/M ratio) altered significantly during treatment of paediatric osteoarticular tuberculosis (OATB) for it to be a useful monitor of the response to therapy. MATERIAL AND METHODS: Thirty immunocompetent paediatric patients with OATB treated with 6 months of uninterrupted multidrug anti-tubercular treatment with resultant clinical and radiological healing of the lesion were included. Haemoglobin, total leucocyte, monocyte and lymphocyte count, and erythrocyte sedimentation rate (ESR) were collected at 0, 2 and 6 months, and were analysed. RESULTS: The L/M ratio altered from high (11) at the start of treatment towards normalisation (9) at 2 months and (7.7) at 6 months. There was no correlation with declining ESR levels. CONCLUSIONS: The L/M ratio may have potential as an effective biomarker response.

Lymphocytic joint fluid in tuberculous arthritis. A review of 30 cases.

UNLABELLED: Several cases of tuberculous arthritis with lymphocytic joint fluid have been reported. This may explain in part the insidious course of tuberculous arthritis. We studied the characteristics of lymphocytic fluid from joints affected with tuberculosis and we looked for specific clinical, radiological, or laboratory test features, comparatively to patients with neutrophilic joint fluid. PATIENTS AND METHODS: We retrospectively reviewed the medical records of 30 patients with tuberculous arthritis, 20 men and 10 women with a mean age of 47.7+/-21.4 years (10-75) and a mean symptom duration at diagnosis of 25.7+/-27.6 months (2-120). RESULTS: Mean joint fluid leukocyte count was 15,181+/-15,635 per mm3 (600-40,000). In joint fluid, neutrophils predominated in 24 patients and lymphocytes in six (20%) patients. Blood cell counts showed no predominance of lymphocytes. No specific clinical, radiological, or laboratory test features were noted in the group with lymphocytic joint fluid. CONCLUSION: We found that a predominance of lymphocytes in joint fluid from patients with tuberculous arthritis was uncommon and was not associated with specific features.

Isolation and characterisation of in vivo released 41 kDa mycobacterial antigen in pulmonary and bone and joint tuberculosis and its identification with H37Ra in vitro released antigen.

OBJECTIVE: To isolate and characterise in vivo released 41 kDa mycobacterial antigen in pulmonary and bone and joint tuberculosis (TB) and its identification with in vitro released ES-41 kDa antigen. DESIGN: Circulating antigen was isolated from confirmed pulmonary tuberculosis serum (PTS) and bone and joint tuberculosis serum (BJS) by trichloroacetic acid precipitation and further fractionation by fast-protein liquid chromatography (FPLC). RESULTS: Fractionation of PTS and BJS by gel filtration column gave six protein fractions each. PTS-G3 and BJS-G3 showed maximum antigenic activity with ELISA. Further fractionation of PTS-G3 and BJS-G3 on cation exchange FPLC gave four different fractions each, of which BJS-G3B was seroreactive similarly to in vitro released 41 kDa antigen (ES-41) isolated from culture medium, whereas PTS-G3C was slightly less seroreactive. BJS-G3B could inhibit binding of in vitro released ES-41 to affinity purified antibodies in inhibition ELISA at lower concentrations than PTS-G3C (2 vs. 20 ng/ml), showing the identical nature of the antigens. Biochemical characterisation showed that circulating antigen PTS-G3C, BJS-G3B and in vitro released ES-41 antigen were lipoproteins in nature. CONCLUSION: This study helped to demonstrate the presence of 41 kDa antigen in the serum of pulmonary and bone and joint TB patients and its identification with H37Ra in vitro released 41 kDa antigen.

[Character of changes in protease inhibitory capacity of blood in experimental osteoarticular tuberculosis]. / Kharakter izmenenii pokazatelei proteazno-ingibitornoi emkosti krovi pri éksperimental'nom kostno-sustavnom tuberkuleze.

All the steps of osteoarticular tuberculosis evolution were accompanied by an increase in proteolytic activity in rabbit blood serum, which was compensated by activation of alpha 1-inhibitor of proteinases at the initial steps of osteitis. The system "proteinases-inhibitors" was imbalanced during the severe phases of arthritis independently on the rate of its development as follows: predominance of proteolytic potential over inhibitory activity, dissimilar activity of alpha 1-inhibitor of proteinases simultaneously with equal rate of alpha 2-macroglobulin content decrease. Dissimilar alterations in the system studied enabled to suggest that differential application of proteinases is suitable for treatment of inflammation with respect to the step of osteoarticular tuberculosis development.

[The epidemiological and clinical aspects of extrapulmonary tuberculosis in children and adolescents in Primorsk Region]. / Epidemiologicheskie i klinicheskie aspekty vnelegochnogo tuberkuleza u detei i podrostkov v Primorskom krae.

The incidence of extrapulmonary tuberculosis has been studied in children and adolescents in the Primorye Territory in 1996 to 2001. There is a decrease in the incidence and prevalence of extrapulmonary tuberculosis in both children and adolescents, which does not reflect the true situation as the process is more frequently diagnosed on their referral in the period of late clinical manifestations. Young age children have fallen ill more frequently (54.7%); 76% of the patients were identified on their referral. Most children are permanent residents and live in the high- and middle-class families. In the clinical pattern, there were predominant ostcoarticular forms of tuberculosis running with a complication of the underlying disease in 80%. The study suggests that there are great gaps in the detection of extrapulmonary tuberculosis in children.

Vertebral osteomyelitis: clinical features and diagnosis.

We aimed to describe clinical and diagnostic features of vertebral osteomyelitis for differential diagnosis and treatment. This is a prospective observational study performed between 2002 and 2012 in Ankara Numune Education and Research Hospital in Ankara, Turkey. All the patients with vertebral osteomyelitis were followed for from 6 months to 3 years. In total, 214 patients were included in the study, 113 out of 214 (53%) were female. Out of 214 patients, 96 (45%) had brucellar vertebral osteomyelitis (BVO), 63 (29%) had tuberculous vertebral osteomyelitis (TVO), and 55 (26%) had pyogenic vertebral osteomyelitis (PVO). Mean number of days between onset of symptoms and establishment of diagnosis was greater with the patients with TVO (266 days) than BVO (115 days) or PVO (151 days, p <0.001). In blood cultures, Brucella spp. were isolated from 35 of 96 BVO patients (35%). Among 55 PVO patients, the aetiological agent was isolated in 11 (20%) patients. For tuberculin skin test >15 mm, sensitivity was 0.66, specificity was 0.97, positive predictive value was 0.89, negative predictive value was 0.88, and receiver operating characteristics area was 0.8. Tuberculous and brucellar vertebral osteomyelitis remained the leading causes of vertebral osteomyelitis with delayed diagnosis. In differential diagnosis of vertebral osteomyelitis, consumption of unpasteurized cheese, dealing with husbandry, sweating, arthralgia, hepatomegaly, elevated alanine transaminase, and lumbar involvement in magnetic resonance imaging were found to be predictors of BVO, thoracic involvement in magnetic resonance imaging and tuberculin skin test > 15 mm were found to be predictors of TVO, and history of spinal surgery and leucocytosis were found to be predictors of PVO.

Tuberculous dactylitis in the setting of low serum vitamin D: a case report.

We present the case of a previously well patient who presented to the Emergency Department of a Dublin hospital with a tuberculous infection of his dominant index finger and a very low serum vitamin D level--this has been implicated in both primary and reactivation infections with Mycobacterium Tuberculosis. This case highlights and reviews both the importance of considering non-endemic pathologies in the setting of a patient base of diverse ethnicity, and the emerging importance of vitamin D in the immune response to M. tuberculosis infection. We discuss the relevant literature to highlight the background of this disease process, and the importance of a multidisciplinary approach to these patients.

Diagnostic usefulness of a T-cell-based assay for osteoarticular tuberculosis.

BACKGROUND: Although diagnosing osteoarticular tuberculosis (TB) remains a challenge, a recently developed Mycobacterium tuberculosis-specific ELISPOT assay for diagnosing TB infection has shown promising results. We performed a prospective, blinded, observational study to compare its diagnostic usefulness with those of conventional tests in patients with suspected osteoarticular TB. METHODS: All patients presenting at a tertiary hospital between April 2008 and September 2009 with suspected osteoarticular TB were enrolled. In addition to conventional tests for TB, we used ELISPOT assays to measure the IFN-gamma response to ESAT-6 and CFP-10 in T-cells in samples of peripheral blood mononuclear cells (PBMC). Patients with suspected osteoarticular TB were classified by diagnostic category. RESULTS: Of the 65 patients with suspected osteoarticular TB, 5 (8%) were excluded due to inconclusive diagnoses. Of the remaining 60 patients, 23 (38%) were classified as having confirmed TB, 3 (5%) as having probable TB, 2 (3%) as having possible TB, and 32 (53%) as not having active TB. Five (8%) patients with probable or possible TB were excluded from the final analysis. Of the 23 patients with confirmed osteoarticular TB, 15 (65%) had TB spondylitis, 4 (17%) had TB arthritis, 2 (9%) had prosthetic joint infection, and 2 (9%) had extra-spinal TB. The sensitivities of the tuberculin skin test (> or =10 mm) and the ELISPOT assay for active osteoarticular TB were 80% (95% confidence interval [CI], 58%-92%) and 100% (95% CI, 85%-100%) (P = 0.04), respectively and their specificities were 68% (95% CI, 51%-81%) and 58% (95% CI, 41%-74%) (P = 0.60), respectively. CONCLUSION: A negative ELISPOT assay using PBMC may be a useful test for excluding a diagnosis of active osteoarticular TB.

[Biochemical criteria of the activity of osteoarticular tuberculosis]. / Biokhimicheskie kriterii aktivnosti kostno-sustavnogo tuberkuleza.

A correlation between the informativeness of the proteinogram, blood ceruloplasmin and haptoglobin levels and connective tissue metabolism indices used to characterize the inflammatory process activity was performed in 55 patients with different stages of tuberculosis of bones and joints. Among the acute phase proteins examined, the most sensitive indices include ceruloplasmin and haptoglobin found in their combination. A higher urinary oxyproline and glycosaminoglycan excretion in different forms of osteoarticular tuberculosis (both of an inflammatory and a degenerative type) should not be interpreted as an indication of activity of the specific process.