| PART?EVALUATION OF ISO-MTB ASSAY FOR RAPID DETECTION OF MYCOBACTERIUM TUBERCULOSIS COMPLEX IN SPUTUM SPECIMENSObjective Isothermal MTB Test (ISO-MTB) is a novel molecular assay for Mycobacteria detection combined recombinase-aid isothermal amplification with flow-through hybridization. We evaluated its capacity to the detection of the Mycobacteria, especially the Mycobacterium tuberculosis complex (MTBC),to explore its further clinical promoting prospect.Methods A total of 270 clinical sputum specimens (117 smear-positive sputum specimens by Auramine O fluorescent stain, and 153 smear-negative sputum specimens) were included in our study; DNA was extracted for ISO-MTB. Culture was regarded as the gold standard to analyze the performance of ISO-MTB in MTBC detection. For comparison, the performance of a conventional real-time PCR assay was analyzed as well, and Chi-square test or Fisher's exact test was utilized to evaluate the difference of the two methods.Sequencing was regarded as the gold standard to analyze the agreement rate of ISO-MTB in Mycobacterium avium complex (MAC) detection. Clinical diagnosis was inquiried to culculate the agreement rate between ISO-MTB and the clinical diagnosis in all the sputum specimens.Results1. ISO-MTB detected two specimens co-infected with MTBC and MAC.An assessment of ISO-MTB was made using the remaining 268 specimens. For MTBC, the overall sensitivity was 92.70% (95%CI, 86.99-96.44), the sensitivity for smear-positive specimens was 95.58% (89.98-98.55) while it was 79.19%(57.85-92.87) for smear-negative specimens. The overall specificity was 96.95%(92.37-99.16), the specificity for smear-negative specimens was 97.67%(93.35-99.52). The overall positive likelihood ratio was 30.36 (11.55-79.77),while the negative likelihood ratio was 0.08 (0.04-0.14). The overall positive predicted value was 96.95% (92.37-99.16), while the negative predicted value was 92.70% (86.99-96.44). No significant differences in MTBC detection results were identified between the PCR assay and ISO-MTB (p>0.05).2. No specimens was detected as MAC alone, but 2 specimens were identified as MTBC-MAC co-infections by ISO-MTB.3. The agreement rate between the clinical diagnosis and ISO-MTB was 94.05%.Conclusion ISO-MTB is an rapid and well-performing method to detect MTBC in clinical sputum specimens, and it could prompt MAC, even co-infected specimens. Thus, ISO-MTB has the potential to be a novel, reliable and rapid method in the clinical identification of MTBC.PART? ASSOCIATION OF INTERLEUKIN-17A AND SUSCEPTIBILITY TO PULMONARY TUBERCULOSIS IN CHINESE HAN POPULATIONObjective We detected gene polymorphisms distribution in the promoter region of the interleukin (IL) -17A gene at position rs4711998 in Chinese Han population, to explore the association of IL-17A rs4711998 single nucleotide polymorphism (SNP) for the susceptibility to pulmonary tuberculosis (PTB).Methods A total of 214 PTB patients and 197 healthy controls in Chinese Han population were included in our study. DNA was extracted from the peripheral blood. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used for genotyping the polymorphisms of IL-17Ars4711998, sequencing was used to verify the accuracy of PCR-RFLP genotyping. Hardy-Weinderg equilibrium was applied to analyze the crowd representativeness of this study. Chi-square test was utilized to evaluate the difference of genotypes and allele frequencies in the case-control populations.To explore the association between IL-17A rs4711998 SNP and PTB susceptibility, logistic regression with adjusted for gender and age was utilized to analyse odds ratio (OR) with 95 % confidence internals (CI).Results1. No significant differences in gender and age composition were observed between case and control groups (p>0.05). The genotype distribution of IL-17A rs4711998 SNP was compatible with the Hardy-Weinderg equilibrium in both case and control groups (p>0.05).2. We detected AA, AG and GG genotypes in IL-17A rs4711998 in both case and control groups, and ino significant difference was observed in the distribution of genotypes and allele frequencies (p>0.05). Compared with AA genotype, neither AG nor GG genotype did not associate with PTB susceptibility(p=0.995;p=0.085).3. When stratified by gender, both in male and female population, no association was observed between any genotype of IL-17A rs4711998 SNP and PTB susceptibility (p>0.05).4. When stratified by age, both in over 40, and less than 40 years old populations, no association was observed between any genotype of IL-17A rs4711998 SNP and PTB susceptibility (p>0.05).Conclusions In the study, we have not found any association between IL-17A rs4711998 SNP and PTB suscetibility in Chinese Han population. |
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