ABSTRACT
Objective To understand the status of healthcare-associated infection(HAI)management in traditional Chinese medicine hospitals as well as integrated traditional Chinese and Western medicine hospitals in Fujian Pro-vince,analyze the existing problems and weak links,and put forward corresponding improvement measures.Methods A questionnaire was designed through literature and expert consultation,from March to April 2016,42 secondary and above traditional Chinese medicine hospitals as well as integrated traditional Chinese and Western medicine hos-pitals in 8 cities of Fujian Province were conducted on-site investigation,data were analyzed.Results A total of 42 hospitals participated in the investigation,92.86% were traditional Chinese medicine hospitals,7.14% were inte-grated traditional Chinese and Western medicine hospitals;all hospitals set up HAI management committees and HAI management groups of clinical departments,there were 100 HAI management professionals(66 were full-time,34 were part-time),nursing staff accounted for 63.00%,junior college and undergraduate personnel accoun-ted for 84.00%,staff with intermediate and senior professional titles accounted for 79.00%.There were significant differences in academic disciplines and education levels among administrators in secondary and tertiary hospitals(P<0.05). All hospitals carried out HAI case surveillance,only 2.38% achieved HAI informational software monito-ring,83.33% carried out comprehensive and targeted monitoring,42.86%,71.43%,and 80.95% of hospitals car-ried out targeted monitoring on multidrug-resistant organisms,surgical site infection,and intensive care unit respec-tively.Conclusion The environment of majority of Chinese medicine hospitals in Fujian Province improved signifi-cantly,organizations of HAI management is rational,staffing and quality of HAI management personnel is imbal-anced,HAI monitoring is still at preliminary stage,lack information management,HAI management in key depart-ments is not optimistic.
ABSTRACT
Objective To establish and optimize a sensitive and specific quantitative realtime polymerase chain reaction(PCR)method for detection of hepatitis B virus covalently closed circular DNA(HBV cccDNA)in liver tissue. Methods Specific primers and probes were designed to detect HBV DNA(tDNA)and cccDNA. A series of plasmids(3.44 × 100-3.44 × 109 copies/μl)containing a full double-stranded copies of HBV genome(genotype C)were used to establish the standard curve of real-time PCR. Liver samples of 33 patients with HBV related hepatocellular carcinoma(HCC), 13 Chronic hepatitis B patients(CHB)and 10 non-HBV patients were collected to verify the sensitivity and specificity of the assay. A fraction of extracted DNA was digested with a Plasmid-Safe ATP-dependent Dnase(PSAD)for HBV cccDNA detection and the remaining was used for tDNA and β-globin detection. The amount(copies/cell)of HBV cccDNA and tDNA were measured by a real-time PCR, using β-globin housekeeping gene as a quantitation standard. Results The standard curves of real-time PCR with a linear range of 3.44 × 100 to 3.44 × 109 copies/μl were established for detecting HBV cccDNA and tDNA, and both of the lowest detection limits of HBV cccDNA and tDNA were 3.44 × 100 copies/μl. The lowest quantitation levels of HBV cccDNA in liver tissues tested in 33 HBV related HCC patients and 13 CHB patients were 0.003 copies/cell and 0.031copies/cell, respectively. HBV cccDNA and tDNA in liver tissue of 10 non-HBV patient appeared to be negative. The true positive rate was increasing through the digestion of HBV DNA by PSAD, and the analytic specificity of cccDNA detection improved by 7.24 × 102 times. Liver tissues of 2 patients were retested 5 times in the PCR for detecting cccDNA and the coefficience of variations on cycle threshold (Ct)were between 0.224%-0.609%. Conclusion A highly sensitive and specific quantitative real time PCR method for the detection of HBV cccDNA in liver tissue was established and could be used for clinical and epidemiological studies.