Preliminary Study On Mycobacterium Tuberculosis Quantitative Distribution In Different Lesions Of Spinal Tuberculosis By Real Time RT-PCR

Objective:To investigate quantitative reverse transcriptase polymerase chain reaction (Real Time RT-PCR) in clinical diagnosis and treatment of spinal tuberculosis in the application of value analysis of Mycobacterium tuberculosis lesions in the spinal lesions of the existence of different circumstances, from the etiological point of view to explore surgical lesion of spinal tuberculosis the scope for the clinical diagnosis of spinal tuberculosis, treatment to help.Methods:20patients with previously untreated patients with spinal tuberculosis, spinal tuberculosis according to CT showed hardened edge to determine whether there is a hardening of all cases, the patients were taking anti-tuberculosis therapy for4weeks after surgery. Intraoperative take all tuberculosis patients with spinal tuberculosis organization, depending on the scope of the invasion of tuberculosis were drawn randomly divided into:group A(caseous necrotic tissue area tuberculosis center), group B (curing tuberculosis edge band area), group C (group B outside the region "sub bone " area) three groups, while group C in accordance with tuberculosis at different distances from the edge of the hardening with "sub-bone" into group C1(3mm hardened band edge out tuberculosis stove tissue area), group C2(C13mm outward edge of the region to organize regional tuberculosis), group C3(group C2area outside edge3mm tuberculosis tissue area), group C4(group C3area outside edge3mm tuberculosis organization area), the distance a (caseous necrotic tissue area tuberculosis center) group C4farthest detected as a result of standardization control. Extracted with Trizol various lesions of spinal tuberculosis Organization RNA, application of quantitative reverse transcriptase polymerase chain reaction (RT-PCR) for Mycobacterium tuberculosis (MTB) in the target gene MPT-64mRNA and reference gene GAPDH were amplified. Statistical analysis was performed using SPSS17.0software, P0.05was considered significant. Results:Real Time RT-PCR detected before each experimental group patients taking INH+RFP+SM, INH+RFP+PZA, INH+RFP+EMB expression of Mycobacterium tuberculosis MPT64mRNA no significant difference between the three groups, each taking a different anti-tuberculosis drugs Low expression levels between the groups group C4, group C2, group C3less the expression of each period. Table5shows that6patients before surgery taking INH+RFP+SM, group A,group B, group C1, mRNA expression levels between the group C2had no significant difference (P>0.05), there are differences between group C2and the other four groups (P<0.05), group A, group B, group C1, the expression of the group is much higher than group C2, C4;7patients before surgery taking INH+RFP+PZA, group A, group B, group C1, group C2no significant difference between them, but each compared with the group C4were significantly different (P<0.01);7patients with preoperative taking INH+RFP+EMB, group A, group B, group C1, group C2expression was significantly higher than the rest of the group C3, C4(P<0.01). Each taking different anti-tuberculosis drugs in the experimental group group C3and C4expression were significantly lower than the rest of the group expression of group (P<0.05). the average reporting time:2days.Conclusions:(1) using Real Time RT-PCR detection of the focus of spinal tuberculosis in Mycobacterium tuberculosis, with rapid,sensitive specificity high;(2) hardening the focus of spinal tuberculosis in the left wall of Mycobacterium tuberculosis with biological activity,complete surgical excision is conducive to the prevention of postoperative recurrence of tuberculosis.(3) from the bacteriological point of view,the successful use of methods of molecular biology research,analysis of the focus of spinal tuberculosis in different tissues(especially the "subhealth bone") the existence of Mycobacterium tuberculosis,survive the situation;(4)"sub-healthy bone tissue " in the group C2(C1area outside edge3mm tuberculosis tissue area) more than the sharp drop in the number of TB, surgery should be based on the specific circumstances of resection combined with image data to determine the scope,so as not to affect the postoperative spinal stability.