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The aim of this study is to develop a rapid and accurately species typing method for Brucella isolates by using High Resolution Melting (HRM ) analysis .Six pairs of primers were used according to the reference for the sequence of pur‐pose gene .Nineteen biotypes of six species Brucella standard strains were identified by PCR‐HRM analysis and this analysis was used to detect the 35 clinical isolates .Results showed Brucella amplified specific melting curves were different from con‐trasted strains with primer Bspp .The six species Brucella standard strains have own characteristic curve shape from each oth‐ers by PCR‐HRM analysis with five pairs of primers .Thirty‐five clinical isolates of Brucella have entirely consistent with PCR‐HRM curve shape with Brucella melitensis standard strains .So ,PCR‐HRM analysis methods can accurately identify Brucella strains ,especially clinical isolated Brucella melitensis ,and may be used in clinical microbiology laboratories .
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Objective To establish a rapid genotyping method of for methicillin-resistant Staphylococcus aureus(MRSA) based on polymerase chain reaction(PCR)-high resolution melting (HRM ) curve analysis and staphylococcal protein A (SPA ) classifica-tion .Methods 71 strains of MRSA clinically isolated were collected as test strains .Gene sequencing and HRM curve analysis were employed to conduct SPA gene typing .Results According to gene sequencing method ,SPA gene of 71 strains of MRSA was divided into four types ,namely t570 ,t030 ,t002 and t588 .The most predominant type was t570 (74 .65% ) ,followed by t030 and t002(both 7 cases) .The result of SPA gene typing by HRM analysis were basically consistent with that by gene sequencing .Con-clusion PCR-HRM analysis is expected to become a fast ,efficient genotyping for MRSA SPA gene ,providing the basis for hospital infection control .
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Objective To evaluated the application value of two kinds of mass spectrometer(MS)and Vitek MS system in the i-dentification of routinely isolated bacteria in clinic.Methods 149 strains of common bacteria(including 14 genera and 30 species)i-solated from blood,urine,cerebral spinal fluid,secretion and sputum samples in our hospital from March 2012 to January 2013 were collected and simultaneously identified by 2 kinds of matrix-assisted laser desorption ionization-time of flight mass spectrometer (MALD-TOF-MS).The identification results were compared with those identified by the conventional biochemical identification (Vitek2 compact).The strains with the inconsistent results identified by 3 kinds of method were confirmed by 16S rDNA gene se-quencing.Results Among 149 common bacteria,the correct identification rates of genus and species by the Bruker Biotyper MS were 98% and 96% respectively and which by the Vitek MS system were 97% and 95% respectively.There were no misidentified bacterial strains by these two kinds of MS.Conclusion No statistical difference in the identification results was observed between these two kinds of MS system(P >0.05).Both exhibit excellent identification level and are suitable for the routine laboratory iden-tification of clinical microorganism.
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Objective To clarify the mechanism of lipoprotein (a) ［Lp (a)］ metabolic disorder in patients receiving maintenance hemodialysis (MHD), and to offer theoretical basis for seeking for modus operandi of disorder control.Methods The apolipoprotein (a)［apo(a)］ isoforms were identified by high-resolution SDS-agarose gel electrophoretic method followed by immunobloting in 61 patients receiving maintenance hemodialysis (MHD group), 51 patients with end-stage renal disease (ESRD group) and 62 healthy controls (healthy control group). The influential factors for elevated serum level of lipoprotein (a) in MHD group were analyzed by applying statistical method.Results No statistical difference of Lp (a) median level was found between MHD group and ESRD group in LWM isoforms, but their level was significantly higher than that of healthy control group. In MHD group, the concentrations of serum creatine(Crea),cystatin C(CysC) and C-reactive protein(CRP) were significantly higher (P<0.05), while the concentrations of albumin(Alb) and Hb were lower than those of healthy control group (P<0.05). The concentration of LDL-C had no significant difference between MHD group and healthy control group. Compared with those of ESRD group, the levels of Crea, CRP and CysC were not significantly increased (P<0.05) in MHD group. There was no difference in the level of Lp (a) between the MHD and the ESRD patients with LMW-apo (a) isoforms (P>0.05), but compared with healthy control group, the serum levels of Lp (a) in the two groups were remarkably increased (P<0.05). The level of Lp (a) with LMW-apo (a) were significantly correlated with Alb and CysC respectively (P<0.05). The level of Lp (a) with HMW-apo (a) of MHD patients were higher than that of ESRD patients (P<0.05). The patients in the two groups had higher levels of Lp (a) as compared with those of healthy control group. The positive correlation of Lp (a) level with CRP and CysC was observed in patients with HMW-apo (a) (P<0.01). As MHD group as concerned, Lp (a) level was associated with Alb and CysC (P<0.01); regression analysis showed that apo (a) isoforms, Alb and CysC were listed in the regression equation, and the determinate coefficient (r2) was 0.348, in which the respective r2 of apo (a) isoforms, Alb and CysC was 0.121, 0.178 and 0.049.Conclusion Multiple factors may contribute to the high level of Lp (a) in MHD patients. Moreover, Lp (a) with different isoforms of apo (a) had different influencing factors. The corresponding therapeutic measure should be taken according to apo (a) isoforms in order to acquire better therapeutic efficacy for MHD patients.
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Objective To investigate the mechanisms of ureaplasma urealyticum(Uu)resistance to macrolide antibiotics.Methods Twenty strains of clinical isolates of Uu with variable resistance to macrolides and reference strain ATCC 27618 were examined for mutations in 23SrRNA.Results As compared with the sequence of reference strain ATCC 27618 and GenBank,three mutations were found in 23SrRNA of Uu clinical isolates.C2243N(TorC)was found in the 23SrRNA in 5 strains with the phenotype resistance to roxithromvcin and azithromycin.A2149C and A2181T were found in the 23SrRNA in 9 strains with the phenotype resistance to roxithromycin and midrange resistance to azithromycin,and 6 strains with the phenotype of sensitivity to roxithromyein and azithromycin.Conclusions The mechanisms of Uu resistance to roxithromycin and azithromycin may be related with the mutations in 23SrRNA.It may warrant further investigation.
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0.05).CONCLUSIONS The rising rates of the drug-resistant Uu strains may be resulted of no-standard treatments.The treatment time will be shortened and the curative effects will be better if the treatments are based on the results of the susceptibility test.
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Objective To observe the relationship between hemoglobin (Hb) and vascular endothelial growth factor (VEGF) in serum of the patients with cancer. Methods Enzyme linked immunosorbent assay (ELISA) was used to detect VEGF levels in serum of the patients with cancers. Results The serum level of VEGF in the cancer group are significantly higher than that in the control group. The serum level of VEGF in patients with low level of hemoglobin ( 123.5 g/L) ( P
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OBJECTIVE To analyze the relationship between the ventilator tube system and VAP.METHODS To investigate the way to use the ventilator tube system more reasonably.RESULTS To replace the ventilator tube every seven days could prevent the VAP effectively and decrease the mechanical ventilation patient′s mortality.CONCLUSIONS Decreasing the VAP incidence is a systematic and integrative work.To reinforce the management of the ventilator tube system is one of the effective methods in preventing VAP.
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Objective To investigate an effective method of breathing apparatus management. Methods According to the defect in process of breathing machine management, breathing apparatus management center was set up, related standards and procedures were established, the staff training was intensified and the management mode was expanded. Results The qualified rate of ventilator sterilization and breathing apparatus function were improved to 100% by applying zero defect quality management. Conclusion It is an effective method to ensure the safety and accuracy in breathing machine manage- ment with applying zero defect quality management.