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Objective To detect dengue virus infection by serological method and to determine the sequences of E gene of dengue virus isolated from Guangzhou in 2006.so as to clarify the possible origin of dengue fever.Methods IgM and IgG antibodies to dengue virus were detected by immunochromatographic test(ICT);NSI antigen and IgM antibody were detected by enzyme-linked immunosorbent assay(ELISA).The virus was cultured and isolated from the serum samples within 2 days using C6/36 cell lines and was identified by immuno-fluorescence assay(IFA)and RT-PCR.The E gene of isolated virus DV1-GZ42/06 was sequenced;homological analysis and phylogenetic tree analysis were performed by comparing with the reference strains and epidemic virus strains.Results The positive rates of IgM and IgG of dengue virus in patients were 89.5%(433/484)and 38.0%(184/484)by ICT,respectively.The positive rates of NS1 antigen were 92.7%(38/41)in day 1 to day 2,83.3%(70/84)in day 3 to day 5,and 10.9%(5/46)in day 6 to day 10;and the IgM detection rates were 2.4%(1/41),51.2%(43/84)and 97.8%(45/46)at the same period by ELISA.Twenty-five strains of dengue virus were isolated from 41 serum samples(6 1.O%)and were identified as type 1 dengue virus by IFA and RT-PCR.The sequencing and phylogenetic analysis of the E gene showed that the homology between the isolated Guangzhou/42/06 strain and standard strain Hawaii/45 was 94.6%.and it had a high homology with the Thailand/NI09V104,Vietnam/06.and Vietnam/07 isolates(99.0%,98.6%and 98.6%,respectively)and belonged to the same cladogram,but had low homology with the isolated strain from Guangdong before 2006.Conclusions The detection of NS1 antigen is important in the early diagnosis of dengue fever.The outbreak of dengue fever in Guangzhou in 2006 was possibly caused by the cases from neighboring countries.
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Objective To investigate in vitro activities of 5 fungal agents(amphotericin B,ketoconazole,fluconasole,5-fluorocytoaine and itroconazole)against Penicillium marneffei,providing references for clinical treatment.Methods E-test was used to test the in vitro susceptibilities of 5 antifungal agents (amphotericin B,ketoconazole,fluconasole,5-fluorocytoaine and itroconazole)against yeast form and mycelial form of 52 Penicillium marneffei strains isolated from our hospital.Results The 90% minimal inhibitory concentrations(MIC90)of amphotericin B,ketoconazole,fluconasole,5-fluorocytoaine and itroconazole against yeast form of Penicillium marneffei were 0.250,0.160,24.000,4.000,0.006 mg/L,the minimal inhibitory concentration(MIC)ranges were 0.004-0.500,0.002-0.016,1.000-256.000,0.002-32.000 and 0.002-0.008 mg/L,respectively;the MIG90 of the 5 agents against mycelial form of Penicil lium marneffei were 1.500,0.125,256.000,24.000 and 0.012 mg/L,respectively;the MIC ranges were 0.064-4.000,0-006-0.940,1-000-256.000,0.125-32.000 and 0.002-0.064 mg/L respectively.Five antifungal agents had different susceptibility patterns against both forms of Penicillium marneffei.And itroconazole was the most susceptible one,ketoconazole follows as the second.There was significant difference between the MICs of the same agent against yeast form of Penicillium marneffei and mycelial form of Penicillium marneffei.Conclusion In vitro antifungal agents suscepbibility tests of Penicillium marneffei can provide important references information for clinical treatment.
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Objective To evaluate bone marrow smear examination in early diagnosis of AIDS complicated with disseminated Penicillium marneffei infection. Methods Seventy-three clinically suspected AIDS patients complicated with disseminated PeniciUium marneffei infection were included in the study. Peripheral blood and bone marrow smear examinations, and the fungal thermally dimorphic culture were performed in all cases. Results PeniciUium marneffei infection was identified in 44 patients by peripheral blood and bone marrow fungal thermally dimorphic culture. The features of the bone marrow smear were as follows : they were all hyperplastic or significantly hyperplastic; there were thickened and increased granules, vacuolization and band-formed in most granulocytes; there were increased and augmented histiocytes, and increased plasma cells. In 12 samples of bone marrow smear, there were phagncytized mulberry-like Penicillium marneffei organisms in the cytoplasm of the histiocytes or the organisms found extracellularly. One sample demonstrated the increased granulocytes and the phagocytized organisms in the neutrophils and monocytes. In 4 samples of peripheral blood smear, there were phagocytized Penicillium marneffe organisms in the neutrophils and monocytes. Conclusion Bone marrow smear examination is of value in early diagnosis of AIDS complicated with disseminated Penicillium marneffei infection, which is 7 to 10 days earlier than routine fungal thermally dimorphic culture.
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Objective To analyze the characteristics of laboratory test resuits of dengue fever(DF)patients in Guangzhou area.Methods Routine tests were performed in the patients admission to hospital. Serology examination was performed in the patients in acute phase or recovery phase.The clinieal symptoms and teatures were analyzed and positive numbers and positivity ratios were calculated.Results The clinical symptoms of the dengue fever were typieal,with the features of fever,headache,myalgia and rash.The leukopenia rate was 76.0%,and the thromboeytopenia rate was 62.6%.The levels of ALT increased in 56.7%patients,and the levels of AST increased in 84.0%patients.Hypopotassemia was found in 46.1%patients.Dengue virus antibody IgM(DF-IgM)was detected positive from the first day to the 16th day of the onset,and the positive rate was 85.9% on the 8th day.Virus loads were positive by fluorescence real-time PCR in seven acute serum samples(within 3 days of the onset)of 51 cases whose DE-IgM were negative all the time, and the results was 105 -106 copies/ml(＜103 copies/ml means negative).Conclusions Clinical manifestations of this DF epidemic were typical including fever,headache,myalgia and skin rash.Most of the patients had decreased leukocyte and thrombocyte obviously.Liver damage was common but kidney damage was seldom.Halt of the patients got hypopotassemia.DF-IgM appeared in very early and persisted for a long time.The detection of DF-IgM within 7 days of the onset was helpful for diagnosis as early as possible.Viral load detected by real-time PCR could be another indicator of early pathogen diagnosis which provides complementation for antibody detection.
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Objective To study the significance of anti-antibody of liver antigens in the diagnosis of autoim- mune hepatitis disease.Methods Patients were divided into three groups according the diseases:autoimmune hepatic disease 45 cases including autoimmune hepatitis(AIH)15 cases,primary biliary cirrhosis(PBC)20 cases,and prima- ry sclerotic cholangitis(PSC)10 cases;Various virus hepatitis 50 cases;Liver damage of unknown cause 30 cases.Au- to-antibody of liver antigens SLA/LP,LKM-1,LC-1,and AMA-M2 were identified by indirect immunofluorescence (IIF)assay and immunoblot assay.Results The positive rates of anti-SLA/LP(46.6%)was significantly higher than those of anti-LKM-1(13.3%),anti-LC-1(0%),and anti-AMA-M2(13.3%)in patients with AIR while these four antibodies were negative in patients with virus hepatitis.The positive rates of anti-AMA-M2 in patients of PBC and unknown liver damage were 95.0% and 6.6%,respectively.Conclusion Anti-SLA/LP is a new specific serum marker in diagnosis of AIR.The auto-antibody detection of liver antigens will be helpful to the diagnosis and therapy of autoimmune hepatic disease.