The Role of Polymerase Chain Reaction (PCR) in Diagnosis of Spine Tuberculosis after Pre-operative Anti-tuberculosis Treatment (original) (raw)
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Multiplex PCR as a novel method in the diagnosis of spinal tuberculosis–a pilot study
Acta Neurochirurgica, 2017
Background Establishment of a reliable and rapid diagnosis is of paramount importance in spinal tuberculosis. The available gadgetry of investigations, such as AFB smear, culture of Mycobacterium tuberculosis, and Uniplex PCR, suffers from a lack of adequate sensitivity and/or a lack of rapidity. Therefore, many times a diagnosis is made either very late in the disease process or sometimes empirical therapy has to be started because a definite diagnosis could not be made. All of these are not ideal situations for a clinician. The present study was done with the aim to establish a rapid and reliable diagnosis of M. tuberculosis infection. This was established by identifying M. tuberculosis genes. Methods The study was done on nine consecutive patients who presented with non-traumatic spontaneous vertebral compression collapse. CT-guided aspirate from the involved vertebra was subjected to Multiplex PCR (MPCR) using three primers: IS6110, protein b, and MPB 64. The aspirate was also subjected to smear and culture. The results of MPCR were compared with the final diagnosis. Results Seven out of nine patients had a final diagnosis of tuberculosis. MPCR was positive in six of these seven patients, thus showing sensitivity of 85.7% and specificity of 100%. Results of MPCR were obtained within 24 h. Conclusions MPCR using IS6110, protein b, and MPB64 primers has a high sensitivity and specificity in rapid diagnosis of spinal tuberculosis. To the best of our knowledge, this has not been attempted before in spinal tuberculosis. This is particularly useful for paucibacillary infections like spinal tuberculosis. However, further studies using large sample sizes are needed to confirm the practical applicability of this technique.
Correlation between PCR and histopathology in TB spine
2020
Introduction: Tuberculosis (Tb) is an infectious disease with enormous mortality and morbidity. In India alone about 33million people are suffering from Mycobacterium tuberculosis and around 3 million are suffering from extra-pulmonary tuberculosis. It is a major burden on health care agency, hence the importance for rapid and accurate diagnosis cannot be ignored. Earlier diagnosis was based on detection of AFB on ZN stains which offers low sensitivity (requires 10^4 bacilli/ml for positive result), Mycobacterium culture is the standard method but due to its long incubation time affects treatment and long term outcome. Histopathological examination provides good information regarding the tissue and sample material but with limited capacity of its specificity the diagnosis and treatment is doubtful, i.e. epithelioid granuloma and caseating necrosis can occur in disease other than Tb. The acknowledgment of ATT therapy was widely used previously to confirm the diagnosis. With the adven...
Microbiological diagnosis of spinal tuberculosis
International Orthopaedics, 2012
Purpose The purpose of this study was to review the clinical features and diagnosis of spinal tuberculosis cases reported in the literature. Methods A medical literature search in the Medline Pubmed database was undertaken to review tuberculosis spinal infection and extra-pulmonary tuberculosis diagnosis improvement. We introduced the following search items and boolean operators: "spinal infection", "spinal tuberculosis infection", "microbiological diagnosis of spinal tuberculosis" and "spinal tuberculosis PCR." Single cases or series without microbiological diagnosis were rejected. Manuscript language was restricted to Spanish, French, and English versions. Results and conclusions Spinal tuberculosis is more common in developing countries and is probably underdiagnosed. Delayed diagnosis is characteristic; it worsens the prognosis and increases morbidity. The microbiological diagnosis is crucial for several reasons. Despite surgical treatment, medical treatment with anti-tuberculous drugs is always necessary. A total of 20-40% of the spinal tuberculosis patients show another locus of infection. Pulmonary location can become a public health problem. Previously treated patients for other tuberculosis locations, incomplete treatments, or poor adherence can change the M. tuberculosis sensitivity pattern. Drug resistance test becomes a major need in the microbiology laboratory. PCR diagnostic techniques advance the diagnosis and increase the sensitivity and specificity rate.
The Role of Polymerase Chain Reaction (PCR) in Diagnosis
Objective: The aim of this study was to evaluate the role of polymerase chain reaction (PCR) in the diagnosis of spinal tuberculosis after 2 weeks of preoperative anti-tuberculosis treatment and to compare PCR to the Löwenstein - Jensen Culture (LJC) and histopathological examination (HPE) methods. Methods: Twenty-five patients were included in this study. Sixteen patients were diagnosed and treated for spinal tuberculosis based on clinical and radiological evidence. Nine patients were controls. The LJC method and HPE of the specimen were performed according to hospital protocol. PCR was performed using primer encoding insertion of sequences IS6110 for mycobacterium tuberculosis complex. Clinical findings and radiological features were the gold standard for comparison. Results: PCR results were 15 positive and one negative. The sensitivity and specificity of PCR was 94% and 100% respectively (with 95% confidence interval [CI] 67% to 99% and 63% to 100%, respectively). HPE results showed 13 were positive and 3 negative in the spinal tuberculosis group; for the control group, all were negative. Sensitivity and specificity value of HPE was 82 % and 100% respectively (with 95% confidence interval [CI] 54% to 95% and 63% to 100%, respectively). Use of LJC showed only one was positive and 15 were negative in the spinal tuberculosis group whole all nine in the control group were negative. Sensitivity and specificity value of LJC was 6% and 100% respectively (with 95% confidence interval [CI] 0.3% to 32% and 63% to 100%, respectively). Conclusion: Our findings showed that the PCR for Mycobacterium tuberculosis is reliable as a method for diagnosis of spinal tuberculosis, even after of 2 weeks of anti-TB treatment, with an overall sensitivity of 94% and specificity of 100%.
Asian Journal of Medical Sciences, 2022
Background: Spinal tuberculosis (TB) is the most common of musculoskeletal TB. The diagnosis of spinal TB is difficult due to the non-specific symptoms. Clinical assessment, along with radiological features and biopsy, plays an important role in early diagnosis of spinal TB. Aims and Objectives: The aims of this study were to analyze clinical, radiological, and pathological features for early diagnosis of spinal TB and to identify other pathological conditions mimicking spinal TB. Materials and Methods: The present prospective study includes 110 patients. All patients were clinically and radiologically examined. The patients underwent percutaneous biopsy of the involved region and tissue samples were subjected to histopathological examination (HPE) and cartridge-based nucleic acid amplification test (CBNAAT) for definitive diagnosis of spinal TB. Results: Out of 110 cases, male preponderance (65.5%) was seen in comparison to females (34.5%). Dorsal spine (53.7%) and lumbar spine (41.3%) were the most common site of involvement. One hundred patients were confirmed as spinal TB by histopathology and molecular diagnosis. Histopathology alone could make diagnosis in 40 cases while molecular diagnosis in 100 cases and 10 cases were non-tuberculous in etiology (metastatic deposits of carcinoma in five cases, pyogenic spondylodiscitis in three cases and primary neoplastic lesion in two cases, one case of giant cell tumor, and one case of hemangioma each). Conclusion: Spinal TB is a deep-seated and paucibacillary condition difficult to diagnose due to inadequate sample. Therefore, multipronged approach by direct smear examination, HPE, and molecular diagnosis is required for early diagnosis. Culture is the gold standard for diagnosis while CBNAAT is highly sensitive and specific that enables rapid detection of tubercular bacilli and should be considered as first-line test.
Journal of Medical Microbiology, 1997
The role of the polymerase chain reaction (PCR) in the diagnosis of tuberculosis in clinical practice remains to be defined; most results have been based on sputum samples. This study systematically compared the relative sensitivity and specificity of a single simplified method for different clinical samples. A wide range of clinical samples, including sputum, bronchoalveolar lavage fluid, cerebrospinal fluid, pleural fluid, gastric aspirate, pus and tissues (both fresh and paraffin-embedded) was tested. This method did not require routine DNA extraction before PCR, and consisted of an optimised single tube PCR amplification designed with different sets of time and temperature profiles. A total of 398 samples from 293 patients was studied. The sensitivity was 100% for all types of specimens, while the specificity ranged from 95% for sputum to 88% for bronchoalveolar lavage fluid and pleural fluid and to 85% for non-pulmonary specimens. This study showed that it was possible to employ a single simplified method with minor modifications for a wide range of specimens in clinical practice without loss of sensitivity and specificity.
Clinical evaluation of a Mycobacterium tuberculosis PCR assay
Journal of clinical microbiology, 1995
On the basis of previously published PCR primer sequences, we have designed a sensitive system for detecting DNA of the Mycobacterium tuberculosis complex (MTB) in patient sputum samples which employs a fast and simplified sample preparation method appropriate for routine diagnostic testing. In order to evaluate the accuracy of the PCR assay, we performed a prospective study with 103 patients, comparing PCR results with culture results of samples obtained from a parallel culture assay as well as with subsequent culture results. Using two MTB-specific PCR primer systems, we found 48 of 49 tuberculosis (Tb) patients to be PCR positive (PCR sensitivity, 0.98). Sixteen of 54 presumably non-Tb patients showed amplifiable MTB DNA (specificity, 0.7). The study demonstrates that for diagnostic applications of MTB PCR two MTB-specific primer pairs should be used. MTB infection is extremely unlikely in cases of MTB PCR-negative samples: with our method for the exclusion of active Tb, the vali...
Journal of Clinical Pathology, 1995
Aim-To investigate the use of the polymerase chain reaction (PCR) in the routine laboratory for the detection of Mycobacterium tuberculosis in clinical samples. Methods-Samples were divided and processed separately for the detection of M tuberculosis by microscopy, culture and PCR. After DNA extraction, PCR was performed with primers specific for the insertion element IS6110 and the product was analysed by agarose gel electrophoresis, Southern blotting or dot blotting and hybridisation with a digoxigenin labelled internal probe. Each sample was tested for inhibitors of Taq polymerase with the aid ofan internal control. Multiple negative and positive controls were used to monitor each step of the procedure. Results-The data from two laboratories, using the same operating procedures, were combined. Of 1957 specimens, 79 (4%) were culture and PCR positive, while 1839 (93.9%) were negative in both tests. Thirty specimens (1-5%) were PCR positive only and nine (0.5%) were culture positive but PCR negative. Conclusion-Using culture and clinical history as the gold standard, sensitivity and specificity for PCR were 92-1% and 99-8%, respectively. With elaborate precautions, PCR is a suitable and reliable method for the detection ofM tuberculosis in clinical samples in a routine microbiology laboratory. (J Clin Pathol 1995;48:810-814)