OBJECTIVE To set-up safe alternatives to ethidium bromide(EB).METHODS Comparative analysis was performed on the DNA staining efficiencies of 4 fluorescent dyes including SYBR Gold,SYBR Green Ⅰ,(GoldView) and EB in preparative agarose gel electrophoresis.RESULTS Although both SYBR Gold and SYBR Green Ⅰ altered electrophoretic mobility and thus DNA size estimates,they were cost-effective alternatives to EB.SYBR Gold was more sensitive than SYBR Green Ⅰ at detecting short fragments,but 50 bp bands were clearly(visible) using either dye when visualized with a long integration time.CONCLUSIONS SYBR Gold or SYBR Green Ⅰ are sensitive and relatively safe alternatives to EB.

OBJECTIVE To develop a real-time fluorescence PCR assay to detect the genes encoding thermolabile(hemolysin)(TLH),thermostable direct hemolysin(TDH) and TDH-related hemolysin(TRH) of Vibrio(parahaemolyticus).METHODS The genes of TDH and TRH were selected as target ones of thermostable direct and TDH-related hemolysin,and TLH gene as a specific genomic marker for V.parahaemolyticus.Designed and synthesized the primers and Taqman probes,we investigated 487 stool samples of doubt foodborne illness patients by real-time fluorescence PCR.RESULTS The sensitivity of the assay for TLH and TDH was 1.0?10~2copies,but the sensitivity of TRH was 1.0?10~3copies. Among 487 samples,112 V.parahaemolyticus strains were found;101 samples of these strains showed the production of TDH;none of them was positive for TRH.CONCLUSIONS The Taqman PCR is a rapid and sensitive method to detect the TLH,TDH and TRH of V.parahaemolyticus,it is well suited for screening large numbers of samples at the same time;and TDH is one of the primary virulence factors in clinical isolated V. parahaemolyticus.

OBJECTIVE To search a method for identifying Clostridium perfringens and genotyping their toxin for gene diagnosis by multiplex PCR.METHODS The mutiplex PCR was developed with three sets of primers(designed) based on the sequences of three C.perfringens toxin genes(CP?,CP? and CPE) published in GenBank to identify C.perfringens and genotype their three toxin genes.RESULTS Three expected(sequences) were (obtained) successfully by multiplex PCR and identified by electrophoresis.CONCLUSIONS The(specific) sequences of C.perfringens could be amplified and their three genes of toxins could be identified by this multiplex PCR(system).Such method should be helpful for developing gene diagnosis well.

OBJECTIVE To analyze three methods with five antifungals susceptibility test.METHODS Eighty(pathogenic) yeasts isolated from various specimens were detected by dish diffusion method for yeast susceptibility testing in three different agars [Shadomy′s modified agar,RPMI 1640 agar,M-H(GMB)agar]with comparison of the Shadomy′s modified agar.RESULTS According to sensibility of five antifungals,RPMI 1640 was the same with Shadomy,but GMB M-H was lower with others.Three different agars had more correlability(P

OBJECTIVE To investigate the distribution of Pseudomonas aeruginosa and its antibiotic resistance in(emergency) intensive care unit(EICU) patients with lower respiratory infections.METHODS The data of the pathogens isolated in sputum from the 25 patients with lower respiratory infections admitted to EICU from(Aug) 2005 to Feb 2006 were collected and analyzed.RESULTS Eighty-one bacteria strains were found in these 25 EICU cases.The Gram-negative bacteria were the main pathogens of infection(43.2%),after them were Gram-positive bacteria(32.1%),and fungi(24.7%).The percentage of P.aeruginosa infection was 13.6% of all(patients).The antibiotic resistance for P.aeruginosa was found to the antibiotics,such as cefotaxime,gentamicin,(SMZ-TMP),minocycline,levofloxacin,and gatifloxacin.CONCLUSIONS The P.aeruginosa infection and its(antibiotic) resistance should be paid more attention in the treatment of the lower respiratory infection patients in EICU.

OBJECTIVE To determine the distribution and drug resistance characteristic of pathogens from(nosocomial) lower respiratory tract infection in wards of respiratory disease and direct clinical doctors to select(antibiotics) correctly.METHODS A total of 159 strains of pathogens from sputum samples of patients with(respiratory) tract infection in wards of respiratory disease in our hospital were identified and the drug resistance tests were performed.In addition,ESBLs-producing G~-bacilli were also detected.RESULTS Among 159 strains from respiratory tract infection,G~-bacilli rated the top one(69.8%),The most common pathogens were Pseudomonas aeruginosa(14.5%),Klebsiella pneumoniae(13.8%),Acinetobacter baumannii(10.7%),(Staphylococcus) aureus(10.7%),and fungi(9.4%).10.8% G~-bacilli produced ESBLs.CONCLUSIONS G~-(bacilli) are the main infectious pathogens in respiratory infection and antibiotic resistance rates continue to(evolve).Therefore bacterial culture and antibiotic susceptibility tests are very important to select antibiotics(correctl)y for treatment of the infections in wards of respiratory disease.

OBJECTIVE To provide reference for prevention and control of nosocomial infection based on the(investigation) of the distribution characteristics of nosocomial infection bacteria and analysis of their antibiotic-(resistance) in wards of gerontologic disease.METHODS The bacteria from specimens of Department of(Gerontology) were cultured and identified,and then antibiotic susceptibility tests were done.These nosocomial(infection) cases judged by the department of nosocomial infection control according to the diagnostic criterion of nosocomial(infection) were analyzed.RESULTS The incidences of nosocomial infection of Department of(Gerontology) were higher in respiratory system,urinary catheter and gastrointestinal tract.A total of 223 strains of pathogens were from(nosocomial) infection and the most commonly encountered pathogens of them were Candida albicans,Pseudomonas aeruginosa,and Escherichia coli.The results of antibiotic susceptibility tests showed that pathogens from nosocomial infection in Department of Gerontology were sensitive to such antibiotics as imipenem,meropenem and polymyxin(98 %), their drug-resistance to the other common antibiotics was very severe.CONCLUSIONS(Possible) measures to reduce nosocomial infection are to strengthen hospital management,reinforce disinfection and sterilization,reasonably use antibiotics,cure the underlying diseases, and shorten(hospital) stay.

OBJECTIVE To study the main bacteria and their drug-resistance of postoperative infections after liver transplantation.METHODS The distribution and drug-resistance profile of 156 strains of bacterial isolates from(various) specimens in 68 liver transplantation inpatients were retrospectively analyzed last year.RESULTS The(incidence) of infections after liver transplantation was 67.6 %.The major bacteria were Klebsilla pneumoniae((17.3%)),Staphylococcus aureus(14.1%),Candida albicans(11.0%),and Pseudomonas aeruginosa((11.0%)).The bacteria had the character of multidrug-resistance and high drug-resistance.The most effective(antibiotics) to Gram-negatives and Gram-positives were still the carbopenems and glycopeptide.CONCLUSIONS(Infection) is a major complication after liver transplantation.Prevention,early diagnosis and treatment of the(infection) are very important.

OBJECTIVE To analyze pathogenic bacterium distribution and antibiotic resistance of our hospital,and provide scientific basis for clinical rational using of antibiotics.METHODS The patients′ clean catch(midstream)(urine) was collected from Jan 2004 to Dec 2005 and cultivated.Antibiotic sensitivity test and adopted by Kirby-Bauer method.RESULTS The pathogenic bacteria mainly consisted of Gram-negatives,among which Escherichia coli was the most frequent,the others in turn were Pseudomonas aeruginosa,Enterobacter cloacae and(Proteus) mirabilis;Enterococcus were the most common among Gram-positives;fungal infection obviously(increased).The bacteria showed different antibiotic resistance rate and multi-drug resistance.CONCLUSIONS It′s very important for making the clinical use of antibiotic more reasonable and controlling drug resistant strains(transmission).

OBJECTIVE To study the constituent ratio of the pathogenic fungi of blood culture in recent 24 months and their resistance in our hospital.METHODS Blood culture of patients in our hospital was performed by BacT/AlerT120 and the isolated pathogenic fungi were identified by API identified tests(API Inc,France).In(addition) antibiotics sensitivity test was by K-B.RESULTS Of the specimens in 4135 cases,there were 110 strains((2.7%)) with Candida albicans(29%).C.tropicalis(21%) and C.portugal(9%).The(specimens) come from(hepatobilliary)(25%),neurosurgery(24%) and emergency(10%) departments.CONCLUSIONS It is important and necessary to monitor the circumstance of fungal(infection) and resistance of the pathogenic fungi due to its(morbidity) increased.