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.

Diagnostic performance of an enzyme-linked immunospot assay for interferon-γ in skeletal tuberculosis.

The aim of this study was to evaluate the diagnostic performance of an enzyme-linked immunospot (ELISPOT) assay for interferon-γ in patients with suspected skeletal tuberculosis (TB). From March 2007 to June 2010, a total of 36 patients with suspected skeletal TB in a tertiary care hospital in Taiwan were enrolled. Twelve patients (35.3%) had culture-confirmed TB, three (8.8%) patients had probable TB, and the remaining 21 (58.3%) patients did not have TB. Fourteen patients with mycobacterial infection had available biopsy or surgical specimens for histopathological examination and 12 (85.7%) specimens had pathological features consistent with mycobacterial infection. Among the 12 patients with positive findings indicating mycobacterial infection, all seven patients with spinal TB and three of five patients with TB arthritis had positive ELISPOT assays. All nine patients with spinal TB had positive ELISPOT assays, but only four of six patients with TB arthritis had positive ELISPOT assays. The sensitivity, specificity, positive predictive value, and negative predictive value for skeletal TB diagnosis by the ELISPOT assay were 86.7%, 61.9%, 61.9%, and 86.7%, respectively. In conclusion, the ELISPOT assay can provide useful support in diagnosing skeletal TB, and spinal TB can be excluded based on a negative ELISPOT assay.

Multicentric tuberculosis at two rare sites in an immunocompetent adult.

The case of a 20-year-old female who presented with refractory coccydynia and sternal pain is described. She was immunocompetent, and had no systemic features. She was diagnosed with tuberculosis of the sternal and coccygeal regions based on magnetic resonance imaging and histopathology of biopsy specimens. Conservative management with oral multidrug antituberculous therapy completely cured the patient, and she had not suffered any recurrence after three years of follow-up. This case highlights the possibility of the multicentric presentation of tuberculosis at two rare sites in the same immunocompetent patient, even though the differential diagnosis was coccydynia.

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.

Cutaneous and articular tuberculosis in a renal transplant recipient.

Seven months after undergoing kidney transplantation, a 56-year-old woman presented with papules and ulcers in her right forearm. The patient received antibiotics for 8 months with limited improvement. Eleven months after symptom onset, she presented with acute arthritis in her left knee. Asynovial fluid culture yielded Mycobacterium tuberculosis, and a forearm ulcer biopsy showed granulomatous inflammation. After surgical fistulectomy and 12 months of tuberculosis treatment, she was cured. Chronic cutaneous ulcers and articular manifestations in TB are rare, but they should always be considered in the differential diagnosis for immunosuppressed patients. Surgical intervention and prolonged treatment might be necessary.

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.

Differential regulation of chemokine secretion in tuberculous and staphylococcal osteomyelitis.

Bone infection or osteomyelitis is characterized by uncontrolled inflammation and destructive bone loss although little is known about immunopathogenesis of infection. We investigated control of chemokine secretion from osteoblasts infected with either Mycobacterium tuberculosis, which normally elicits a granulomatous host response, or Staphylococcus aureus, which drives a host response dominated by neutrophil influx. We show that M. tuberculosis infection of cultured and primary osteoblasts induces extensive secretion of the chemokines interleukin (IL)-8, inducible protein (IP) 10, RANTES, and monocyte chemoattractant protein (MCP) 1 within 72 h (1630 +/- 280 pg/ml per 4 x 10(5) cells, 74,130 +/- 8480 pg/ml per 4 x 10(5) cells, 18,330 +/- 3040 pg/ml per 4 x 10(5) cells, and 138,670 +/- 13,340 pg/ml per 4 x 10(5) cells, respectively, for MG-63 osteoblasts). S. aureus infection also results in secretion of these chemokines but secretion is delayed and of lesser magnitude (210 +/- 10 pg/ml per 4 x 10(5) cells, 11,570 +/- 1240 pg/ml per 4 x 10(5) cells, 930 +/- 34 pg/ml per 4 x 10(5) cells, and 13,770 +/- 720 pg/ml per 4 x 10(5) cells for IL-8, IP-10, RANTES, and MCP-1, respectively). The minimal up-regulation of secretion of the neutrophil attractant IL-8 in staphylococcal infection is both striking and unexpected. In both infections, chemokine secretion was dependent on the presence of live organisms. Differences in kinetics and magnitude of chemokine secretion are associated with distinct patterns of mRNA expression, as assessed by ribonuclease protection assay (RPA) and reverse-transcription polymerase chain reaction (RT-PCR). In addition, nuclear localization of the transcription factor activator protein (AP) 1 in M. tuberculosis-infected osteoblasts also is distinct as compared with S. aureus-infected cells. In summary, this study shows that osteoblasts have an important pathogen-specific role in control of chemokine gene expression and secretion during the human immune response to osteomyelitis.

Serological tests in the differentiation of staphylococcal and tuberculous bone disease.

The haemagglutination test for antileucocidin is frequently positive in cases of bone tuberculosis in the absence of obvious staphylococcal infection. This test is therefore of little practical use in the differentiation of staphylococcal and tuberculous bone disease, and its use has been discontinued at the Royal National Orthopaedic Hospital. The antigamma haemolysin test in bone tuberculosis appears to give rise to few false positive results. Our observations confirm that the anti-alpha haemolysin and antigamma haemolysin tests used together reveal about 80 percent of cases of staphylococcal bone infection on first presentation or relapse.

Humoral immune response against 38-kDa and 16-kDa mycobacterial antigens in bone and joint tuberculosis.

SETTING: The diagnosis of bone and joint tuberculosis (BTB) is difficult, and diagnostic delays often occur. A reliable serological test detecting anti-mycobacterial antibodies would thus be of some use in this form of the disease. OBJECTIVE: To evaluate the diagnostic accuracy of an assay detecting IgG against 38-kDa and 16-kDa recombinant mycobacterial antigens in BTB. MATERIALS AND METHODS: In a prospective study, serum samples from 124 subjects were examined: 30 BTB cases, 40 non-specific bone and joint infection patients (NSBI), 30 lung cancer patients (LC), and 24 healthy volunteers (HC). An ELISA-based test (Pathozyme TB complex plus) was used. RESULTS: The cut-off level was established at 150 U/ml according to receiver operating characteristic (ROC) curves. The quantified level of sensitivity of the test detecting BTB was 56%, at a specificity of 99%. The positive and negative predictive values were respectively 94% and 88%. Mean IgG level in the BTB group was 470 +/- 761 U/ml (mean +/- SD), and was significantly higher than the antibody level in the control groups (NSBI 58 +/- 42 U/ml, LC 43 +/- 38 U/ml, HC 40 +/- 29 U/ml). CONCLUSION: The test presents an acceptable level of sensitivity and very good specificity in the diagnosis of BTB, and can be used in combination with other methods to increase diagnostic accuracy in this disease.

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.

[Detection of serum antibody against lipoarabinomannan-B in Mycobacterium tuberculosis (H37Ra)].

The serum antibody to lipoarabinomannan-B(LAM-B) purified from mycobacterium tuberculosis (H37Ra) was tested by ELISA in 250 sera, including sera from patients as follows: tuberculosis 96, tubercular pleurisy 11, renal tuberculosis 2, bone and joint tuberculosis 33, tubercular meningitis 16, pulmonary cancer 22, leprosy 20 and normal subjects 50. The positive rate of pulmonary tuberculosis is 69.8%, which is of a similar extent in sera from patients with tuberculosis of miscellaneous organs to be tested except tubercular meningitis, in which only 18.8% positive rate was observed, indicating the blockage of antibody releasing from pathologic foci into blood stream by blood-brain barrier. The positive rates of leprosy and normal subjects are 50.0% and 2.0% respectively. No antibody was found among 22 patients with pulmonary cancer. It is suggested that the existence of an active tubercular lesion in the host might be the basic prerequisite for a positive LAM-B antibody detection. Although LAM-B is a common antigen of both mycobacterium tuberculosis and mycobacterium leprae, the low prevalence of leprosy in China makes little influence of the practicability of using this ELISA in epidemiological study and in clinic as a adjutant tool for tuberculosis diagnosis and differential diagnosis.

Wrist tuberculosis in cladribine--induced remission of hairy cell leukosis.

A 45-year-old man with "hand-shoulder" syndrome developing eight-months after Cladribine-induced remission of an 11-year-old hairy cell leukosis is presented. Wrist bone biopsy was performed because of failure of the algodystrophy treatment and radiographic findings of progressive osteoporosis. Caseating epiteloid granulomas abundant in Langhans cells were found histologically and later Mycobacterium tuberculosis species was isolated in culture specimen. Fistulas were formed that healed after a prolonged anti-tuberculosis therapy. The role of cellular immunity deficiency in Cladribine-treated hairy cell leukosis that predisposes to mycobacterial infection is discussed.

[The immunological aspects of a hip joint lesion in childhood]. / Immunologicheskie aspekty porazheniia tazobedrennogo sostava v detskom vozraste.

The paper presents an in-depth analysis of immunological mechanisms of the hip joint synovitis development in children. Studied in the pediatric patients with transient coxitis, Perthes disease, and tuberculous coxitis (n = 54, 15, and 15 respectively) with the aid of flow cytofluorometer FACStar PLUS was the subpopulation of immunocompetent cells with a wide spectrum of MCAB. Controls (n = 35) were sex and age-matched. Comparative analysis of immunological parameters in the above children disclosed a toxic-and-allergic origination of transient synovitis with predominant affection of the T-cell link of immunity (T-helpers failure and activation of T-suppressors). Joint mucose injury was noted to be developing against the background of reduction of serum IgA. Activation of non-specific mechanisms (natural killers, increase in SDH activity) is of compensatory character. Mechanisms of activation of cell processes (expression of panmitogenic receptor CD4+ antigen) in Perthes disease warrant further studies. In patients with tuberculous coxitis, immunological incompetence develops in the presence of cytotoxic reactions and activation of the B-link, with the production of IgM being on the increase but with no essential changes in the IgG fraction. It is necessary that selective stimulators of T-helpers be included into the treatment scheme together with drugs capable of exerting a selective effect on the T-link of immunity.

Interleukin-3 and interleukin-17 do not play a dynamic role in the immunopathogenesis of osteoarticular tuberculosis.

BACKGROUND: Osteoarticular tuberculosis accounts for one to three per cent of all cases of active TB. IL-3 stimulates the proliferation, differentiation and survival of pluripotent stem cells. IL-17 has shown to promote inflammatory cell recruitment and granuloma organization throughout infection with Mycobacterium tuberculosis. During the chronic phase of the infection, a balance between Th1 and Th17 responses needs to be achieved to limit immunopathology. AIM: To correlate the serum levels of IL-3 and IL-17 at presentation and after completion of treatment in clinicoradiologically proven cases of osteoarticular tuberculosis. METHODS: 32 clinicoradiologically confirmed cases of osteoarticular tuberculosis were included. Archived serum samples of eight patients of osteoarticular tuberculosis of an earlier study, confirmed by PCR, AFB smear or by histopathology with previously determined IL-12 and TGF-beta levels were available. A detailed history was noted and their general physical, local and relevant systemic examination was performed. Various laboratory parameters including TL-3 and IL-17 levels in serum were estimated at presentation and at six months of DOTS CAT-1 treatment. RESULTS: There was a significant improvement in the clinical and radiological parameters after treatment. No correlation was found between IL-3 and IL-17 levels before and after treatment. A significant correlation (p value= 0.022) was shown between levels of IL-3 and IL-12 after six months of treatment. CONCLUSIONS: Qualitative and quantitative fluctuations in IL-3 and IL-17 levels were not able to serve as useful indices of disease activity.

Cutaneous and articular tuberculosis in a renal transplant recipient

Abstract Seven months after undergoing kidney transplantation, a 56-year-old woman presented with papules and ulcers in her right forearm. The patient received antibiotics for 8 months with limited improvement. Eleven months after symptom onset, she presented with acute arthritis in her left knee. Asynovial fluid culture yielded Mycobacterium tuberculosis, and a forearm ulcer biopsy showed granulomatous inflammation. After surgical fistulectomy and 12 months of tuberculosis treatment, she was cured. Chronic cutaneous ulcers and articular manifestations in TB are rare, but they should always be considered in the differential diagnosis for immunosuppressed patients. Surgical intervention and prolonged treatment might be necessary.

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.

Multifocal osteoarticular tuberculosis in children.

PURPOSE: To review records of 16 children with multifocal osteoarticular tuberculosis. METHODS: Records of 7 girls and 9 boys aged one to 14 (mean, 6) years with multifocal osteoarticular tuberculosis were reviewed. Haematological tests and radiographs of the chest, whole spine, pelvis, knees, elbows, hands, and feet were taken. The diagnosis was confirmed histologically. Patients were treated with standard 4-drug antitubercular chemotherapy (isoniazid, rifampicin, ethambutol, pyrazinamide) for 2 months, followed by a 2-drug regimen (isoniazid and rifampicin) for 10 months. Supportive treatment (deworming and nutritional advice) was also provided. RESULTS: All 16 patients were immunocompetent. Pain and swelling around the lesions were the main symptoms; fever was not common (2 cases only). No patient reported weight loss or night sweats. The mean number of bony lesions was 3.4 (range, 2-15) per patient. Appendicular (hands and feet) involvement was more common than axial (spinal) involvement. Radiological appearances of the lesions were cystic, irregular, lytic, and with or without sequestrum/ periosteal reaction. Some lesions were asymptomatic and detected incidentally on radiographs. Only one patient had active chest lesions. Five patients had spinal involvement but no neurological deficit. No patient underwent any surgical intervention, except for diagnostic biopsy. The mean follow-up period was 18 (range, 6-24) months. All patients showed complete healing within one year of chemotherapy. There were residual deformities and restriction of joint movement in patients with advanced articular and axial osteoarticular involvement. CONCLUSION: Children with multifocal osteoarticular tuberculosis were usually immunocompetent. Appendicular involvement was common, but concomitant chest involvement was uncommon. Standard multidrug antitubercular therapy and nutritional supplementation achieved good outcome.