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Objective To optimize the experimental system of dual molecular beacon to rapidly detect My-cobacterium tuberculosis and its resistant strains.Methods Fluorescence quantitative PCR was carried out by selecting different magnesium ion concentration,annealing temperature and primer concentration respectively. Finally,the optimum reaction conditions were obtained.Results In order to ensure the efficiency of amplifica-tion and no non-specific amplification,the final selection of the best conditions were as follows,the concentra-tion of Mg2+was 3.0 mmol/L,annealing temperature was 60 ℃,and the concentration of primers was 0.3 mmol/L.Conclusion The optimal condition of dual molecular beacon experiment was established,which en-sured that the detection of Mycobacterium tuberculosis by molecular beacon quantitative PCR had the advanta-ges,such as simple operation,rapid speed,high sensitivity(the minimum detection limit was 1 CFU/mL)and specificity(only Mycobacterium tuberculosis complex including drug-resistant strains could be detected),good reproducibility(coefficient of variation was < 5%)and other advantages.The study provides the necessary conditions for the dual molecular beacon detection of Mycobacterium tuberculosis.
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Objective To investigate season distribution,biological typing and drug resistant of Haemophitus in Qingdao Central Hospital.Methods The sputum and throat swab were collected from patients with respiratory tract infection,221 Haemophilus strains were identified and typed by the manual method and MicSCAN4 automatic analyzer,HNID identification plate.Antimicrobial susceptibility was tested by Kirby-Bauer method,and cephalosporins nitrate thiophene paper method was used to detect β-lacta-mase.Results A total of 96 strains of Haemophilus influenzae(1.6%)were isolated,10(10.4%)strains of Haemophilus influenzae were identified as type Ⅰ,31(32.3%)as type Ⅱ,40(41.7%)as typeⅢand 1 5(1 5.6%)as other types.A total of 125 strains Hae-mophitus parl influenzae(2.1%)were isolated,1 5 (12.0%)strains of Haemophilus parl influenzae were identified as type Ⅰ,23 (18.4%)as typeⅡ,69(55.2%)as type Ⅲ and 18(14.4%)as type Ⅳ,other types were not identified.The highest infected rate was in winter.Resistance rate of Haemophilus influenzae and Haemophitus parl influenzae to ampicillin were 40.6% and 44.8%,to tri-methoprim-sulfamethoxazole were 5 1.0% and 66.4%.The prevalence ofβ-lactamase of all strains were 40.6%and 44.8%.But sus-ceptible rates of Haemophilus to cefotaxime,cefuroxime,meropenem,chloramphenicol were over 90.0%.Conclusion The respira-tory tract infections to Haemophilus influenzae and Haemophitus parl influenzae is more frequently found in winter.Type Ⅱ and type Ⅲ are the most prevalent types.The resistance rates of Haemophilus to ampicillin and trimethoprim-sulfamethoxazole are in-creasing,should not be used as empirical treatment of Haemophilus infection.Antibiotics such as cefotaxime,cefuroxime,meropen-em could be chosen for the treatment of respiratory tract infection caused by Haemophilus.
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Objective To compare the performance of high‐throughput ELISA and ECLIA in the determination of carcinoembry‐onic antigen (CEA) .Methods The CEA concentration of serum samples were respectively determinate by high‐throughput ELISA and ECLIA ,and the results were compared .Results Two kinds of detection methods could both accurately reflect the concentration of serum CEA .There was no significant difference between the results of two methods (P> 0 .05) .Within the linear range ,the CEA result of high‐throughput was correlate closely with that of ECLIA (r=0 .922 8 ,P<0 .01) .Conclusion High‐throughput ELISA can accurately detect the serum CEA concentration .
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Objective To monitor the resistance of Streptococcus pneumoniae following the 2004 in Qingdao area, and to provide a reasonable basis for clinical application of antimicrobial agents. Methods Collecting respiratory tract, blood, cerebrospinal fluid and other specimens from out-patient and in-patients of some hospital in Qingdao from January 2005 to December 2008. According to the recommendation of NCCLS, antibiotic resistance analysis of 11 kinds of antibiotic to the isolated 231 Streptococcus pneumoniae by micro-agar dilution method, and analysis resistance trends and age differences. Results The results showed that the rate of Streptococcus pneumoniae not sensitive to penicillin is 23. 38% (PRSP: 9.52% , PISP: 13. 85% ) , resistant to cefotaxime is 9. 96% (23/231), resistant to amoxicillin is 12. 55% , resistant to erythromycin is 90. 48% (209/231). PRSP rate of patients younger than 14 years of age 27. 91% (12/43), significantly higher than that of the PRSP rate of adults 5. 38% (10/186). Conclusion The rate of resistant to penicillin Streptococcus pneumoniae increased significantly from 2004, and an increasing trend year by year, the resistance of Streptococcus pneumoniae is also a rising trend year by year. For patients infected low penicillin-resistant Streptococcus pneumoniae in this region, cefotaxime, amoxicillin are preferred drugs.