Prevalence and distribution of hepatitis C among high-risk populations in Pudong New Area, Shanghai City / 中华传染病杂志

Objective:To investigate the prevalence of positive hepatitis C virus (HCV) antibody, HCV RNA and genotype distribution of HCV in high-risk populations in Pudong New Area, Shanghai City, so that to provide evidence for making "hepatitis C micro elimination" strategies in Shanghai area.Methods:A survey with proportional sampling method was conducted among the high-risk populations, including people who inject drugs (PWID), voluntarily or compulsorily accepted drug detoxification or methadone treatment, human immunodeficiency virus voluntary counseling and testing (HIV VCT) outpatients, sexually transmitted disease (STD) outpatients, and commercial sex workers, who participated in the routine physical examination activities held by the community health service centers and public hospitals of Pudong New Area from July 2021 to November 2022. The residual plasma samples were collected from medical examinations. HCV antibody was tested in all samples. HCV RNA and HCV genotype were tested in samples with positive HCV antibody results.Results:A total of 1 000 HCV high-risk people were screened, including 453 PWID, 166 human immunodeficiency virus (HIV) infectors, 245 STD outpatients, and 136 commercial sex workers. The positive rates of HCV antibody in the four categories of personnel were 21.85%(99/453), 1.81%(3/166), 1.22%(3/245) and 0(0/136), respectively. The positive rate of HCV RNA was 42.68%(35/82) among HCV antibody positive people in high-risk populations. As much as 74.29%(26/35) of HCV RNA positive people had junior high school education or less, and 77.14%(27/35) of them were not married. Among the 12 samples tested for HCV genotype, five were genotype 3, five were genotype 6, and two were subtype 1b.Conclusions:PWID is the main high-risk HCV infection population, who should be the target of the following "hepatitis C micro elimination" strategies. The proportions of genotype 3 and genotype 6 are high in the high-risk HCV infection populations, and the pan-genotype direct-acting antiviral agent treatment may be more suitable in this situation. HCV infected persons in high-risk groups have low education level and marriage rate, which indicates that education and care in community are needed.

Therapeutic effects of dialectical addition and subtraction treatment based on self-made Qufeng Zhike Decoction on cough caused by wind dryness invading the lung / 中国基层医药

Objective:To investigate the therapeutic effects of dialectical addition and subtraction treatment based on self-made Qufeng Zhike Decoction on cough caused by wind dryness invading the lung. Methods:Thirty patients with cough after catching a cold who were admitted by Huainan Hospital of Traditional Chinese Medicine from May 2021 to May 2022 were included in this study. These patients were treated with self-made Qufeng Zhike Decoction (first decocting the ochre for 30 minutes, then decocting inula flower in cloth bags, or traditional Chinese medicine drug granules for drinking with boiled warm water), 1 dose per day, 5 doses in total at first diagnosis. During the follow-up visit, patients were given an dialectical addition and subtraction treatment according to the improvement of cough. Five days of dialectical addition and subtraction treatment were taken as one course of treatment, and two or three courses of treatment were used. Therapeutic effects and adverse reactions were recorded. Scores of cough, expectoration and pharyngeal itching before and 15 days after treatment were recorded. Results:All 30 patients actively cooperated with the treatment. Among 30 patients, treatment was remarkably effective in 19 patients, effective in 9 patients, and ineffective in 2 patients, with a total response rate of 93.33% (28/30). Among 30 patients, two complained of mild stomach discomfort (relieved after taking medicine 1 hour after a meal). No obvious adverse reactions were found in the remaining patients. Compared with before treatment, scores of cough, expectoration and pharyngeal itching were significantly decreased after treatment [cough: (2.38 ± 0.51) points vs. (1.00 ± 0.85) points; expectoration: (0.88 ± 0.54) points vs. (0.29 ± 0.08) points; pharyngeal itching: (0.98 ± 0.67) points vs. (0.65 ± 0.33) points, t = 2.36, 2.98, 2.01, all P < 0.05]. Conclusion:Dialectical addition and subtraction treatment based on self-made Qufeng Zhike Decoction for treatment of cough caused by wind dryness invading the lung has a promising curative effect, without obvious adverse reactions.

Clinical characteristics of patients with Klebsiella pneumoniae pyogenic liver abscess / 中华内科杂志

Objective:To summarize the clinical features of patients with Klebsiella pneumoniae pyogenic liver abscess(KP-PLA). Methods:Clinical data of 133 patients with pyogenic liver abscess(PLA) and positive results of blood or pus culture were retrospectively analyzed in Huashan Hospital Affiliated to Fudan University from 2009 to 2018. According to the culture results, patients were divided into KP-PLA group ( n=92) and non-KP-PLA group ( n=41). Results:KP-PLA and non-KP-PLA were similar in gender composition with males accounting for 67.39% and 70.73%, and had age of (56.8±13.8) years and (55.0±13.0) years (χ 2=0.146, 0.708, P>0.05) respectively. The underlying diseases were more common in KP-PLA group, including diabetes accounting for 45.65% and 24.39%, and hypertension accounting for 32.61% and 14.63% (χ 2=5.384, 4.642, P<0.05) respectively. Patients with KP-PLA had more invasive infections beyond liver than those with non-KP-PLA, which were 27.17% and 9.76% (χ 2=5.046, P=0.025). The laboratory results showed that hemoglobin levels in KP-PLA and non-KP-PLA were (109.88±20.97) g/L and (97.75±20.25) g/L ( t=3.086, P=0.002). Serum alkaline phosphatase levels were 146.50 (114.50, 237.50) U/L and 220.50 (120.00, 316.75) U/L in KP-PLA and non-KP-PLA ( U=2 239.500, P=0.048) patients. Conclusions:KP-PLA mainly develops in middle-aged and elderly men, especially those with diabetes and hypertension. Patients with KP-PLA need to be paid more attention for invasive manifestations beyond liver.

Analysis of the changes and efficacy of antiviral treatment regimens in patients with chronic hepatitis C from 2014 to 2019 / 中华传染病杂志

Objective:To analyze the changes and efficacy of antiviral treatment regimens in patients with chronic hepatitis C.Methods:This was a single center retrospective study. A total of 157 patients with chronic hepatitis C in Huashan Hospital, Fudan University from January 2014 to February 2019 were included. Clinical informations of antiviral treatment and follow-up were collected. The sustained virologic response (SVR) rate and adverse events in patients receiving different antiviral regimens were compared. Chi-square test was used for statistical analysis.Results:Among the 157 patients, 133 patients had sufficient follow-up data. Seventy-one patients received treatment before 2017, among which 63 patients received interferon regimens and the SVR rate was 74.65%(53/71). Sixty-two patients received treatment after 2017, among which 61 patients received direct-acting antiviral agents (DAA) regimens and the SVR rate was 98.39%(61/62). The difference in SVR rate between the two groups was statistically significant ( χ2=15.230, P<0.01). In 69 patients who received DAA regimens from 2014 to 2019, the SVR at post-treatment week 12 (SVR12) was 95.65%(66/69). Among 43 patients who received DAA regimens containing sofosbuvir, the SVR12 rates of patients with hepatitis C virus genotype 1, 3 and other genotypes were 15/15, 5/6 and 90.91%(20/22), respectively. All the 26 patients who received DAA regimens non-containing sofosbuvir achieved SVR12. The SVR12 rates of patients with different hepatitis C virus genotypes and DAA regimens were not significantly different ( χ2=5.243, P=0.263). The incidences of adverse events in pre-2017 group and post-2017 group were 84.62%(77/91) and 6.06% (4/66), respectively. The difference was statistically significant ( χ2=94.520, P<0.01). The most common adverse events were decreases in neutrophil cell count, decreases in hemoglobin level and decreases in platelet count. Treatment was ceased in six patients due to adverse events. Conclusions:After 2017, the majority of patients with chronic hepatitis C received DAA regimens instead of interferon regimens. The SVR rate increases and the incidence of adverse events decreases along with the changes of leading treatment regimens.The SVR12 rate is higher in patients receiving DAA regimens, regardless of hepatitis C virus genotypes.

Clinical observation of the therapeutic effect of Xiaochaihu decoction addition and subtraction on cough variant asthma based on the theory of " Shaoyang as the pivot" / 中国基层医药

Objective:To explore the therapeutic effect of Xiaochaihu decoction addition and subtraction on cough variant asthma(CAV) based on the theory of " Shaoyang as the pivot" .Methods:From May 2018 to May 2019, 23 patients with CAV were selected in the Traditional Chinese Medicine Hospital of Huainan, all of whom were diagnosed as CAV by bronchodilation test.The patients were first treated with Xiaochaihu decoction addition and subtraction, 7 doses, decocting in water, oral, 1 dose per day.Add and subtract the prescriptions according to the changes of the condition.The curative effect and adverse reactions were observed, and the recurrence was observed for half a year.Results:Of 23 patients, 11 cases were obviously effective, 9 cases were effective and 3 cases were ineffective, the total effective rate was 87.0%(20/23). Course of treatment: of the 11 cases with obvious effect, 4 cases were treated for 3 weeks, 5 cases for 4 weeks, 2 cases for 5 weeks, and 9 cases with effective effect were treated for 5 weeks.It was suggested to use other treatment for 3 cases with ineffective effect after 6 weeks of treatment.No obvious adverse reactions were found in 22 patients, one patient had slight nausea, which was relieved by himself.All 23 cases were followed up for half a year, 3 cases recurred, the recurrence rate was 13.0%(3/23).Conclusion:Based on the theory of " Shaoyang as the pivot" , the treatment of CAV with Xiaochaihu decoction addition and subtraction has high clinical effective rate, low recurrence rate and no obvious adverse reactions.

Clinical significance of nontuberculous mycobacteria isolated from respiratory specimens / 中华传染病杂志

Objective@#To determine the clinical significance of nontuberculous mycobacteria (NTM) isolated from respiratory specimens. @*Methods@#Clinical data of patients with NTM strains isolated from the respiratory tract between January 2014 and February 2017 were retrospectively analyzed. Clinical significance of NTM isolated strains was evaluated based on diagnostic criteria of NTM pulmonary diseases from American Thoracic Society (ATS). Quantitative data of two groups were analyzed by independent t test. Categorical data were analyzed by Pearson χ2 test or Fisher exact test. @*Results@#Totally 352 NTM strains from 257 patients had been isolated between January 2014 and February 2017. Among 15 identified NTM species, M. intracellular (51.7%, 182/352), M. abscessus (25.6%, 90/352), and M. avium (10.8%, 38/352) were predominant. Of the 157 patients with full clinical data involved in the analysis, 58 (36.9%) patients were determined to have definite disease, and 34 patients (21.7%) were designated as probable disease candidates, and 16 (10.2%) patients were regarded as uncertain disease, and 49(31.2%) patients were diagnosed as unlikely disease. The age of 58 patients with definite disease was (63.9±12.7) years, and 48.3% (28/58) were female. M. intracellulare (55.2%, 32/58) was the main cause of pulmonary NTM disease, followed by M. abscessus (25.9%, 15/58) and M. avium (12.1%, 7/58), while other NTM species only accounted for 6.8% (4/58). Definite cases with M. intracellulare, M. abscessus, M. avium, M. kansasii, M. marseillense, and M. columbia accounted for 35.2% (32/91), 57.7% (15/26), 7/15, 2/3, 1/3 and 1/2, respectively, among patients with corresponding isolations, while patients with other species of isolation did not meet the diagnostic criteria. Patients with clinical significant isolation of NTM were older than those without clinical significance (χ2=3.603, P=0.000), and proportion of anti-acid staining positivity of patients with clinical significance was higher than that of patients without clinical significance (χ2=18.815, P=0.000). The proportion of M. abscessus in patients with clinical significance was higher than that in patients without clinical significance (χ2=6.313, P=0.012). However, there was no significant difference between the two groups in the isolation of M. Gordon (Fisher exact test, P=0.028). The proportion of M. abscessus lung disease in women was 11/15, which was higher than that of M. intracellulare lung disease (41.5%, 17/41), and the difference between the two groups was statistically significant (χ2=4.462, P=0.035). There was no significant difference of clinical symptoms and underlying diseases in NTM lung disease among different groups (all P>0.05). In these patients with definite disease, 39.7% (23/58) of them manifested the upper lobe cavitary form, 53.4% (31/58) exhibited nodular bronchiectatic form, and only 6.9% (4/58) exhibited unclassified form. The upper lobe cavitary form (43.8%, 14/32) and the nodular bronchiectatic form (53.1%, 17/32) dominated in patients with M. intracellulare lung disease. M. abscessus lung disease was dominated by the nodular bronchiectatic form (11/15) while the upper lobe cavitary form only accounts for 3/15. There was no significant difference of image characteristics between NTM lung disease and other different groups (all P>0.05). @*Conclusions@#36.9% of the patients with NTM isolates met the ATS diagnostic criteria for NTM lung disease. Different species have different clinical significance. M. intracellulare and M. abscessus are the most predominant NTM isolated species that cause NTM lung disease. Majority of patients manifest as the upper lobe cavitary form, followed by the nodular bronchiectatic form.

Clinical retrospective analysis of Klebsiella pneumoniae bloodstream infection / 中华传染病杂志

Objective To explore the clinical characteristics,drug resistance and prognosis of Klebsiella pneumoniae bloodstream infection (KP-BSI),and to analyze the risk factors of death and drug resistance.Methods The clinical data of hospitalized patients with KP-BSI from April 2015 to April 2017 in Huashan Hospital were retrospectively analyzed.Continuous variables were compared using t test.Categorical variables were compared using x2 test or Fisher exact test.The independent risk factors for death were determined by logistic regression model.Results The majority of the 74 patients with KP-BSI were male (67.6％) and elderly patients (78.4％).Nosocomial infection occurred in 58 cases (78.4％) and a total of 24 (32.4％) cases died.The patients were widely distributed in various departments of the hospital.The first was the Department of Infectious Diseases (29.7％),followed by the intensive care unit (23.0％).The patients were often complicated with various underlying diseases and the most common was pulmonary infection (56.8％).There were 45 (60.8％) multiple drug resistance (MDR) strains and 29 (39.2％) Carbapenems resistant Klebsiella pneumoniae (CRKP) strains.There were significant differences of nosocomial infections (x2 =4.655,P =0.031),deep venous catheters (x2 =5.432,P-0.02),and invasive mechanical ventilation (x2 =7.630,P =0.006) between MDR and non-MDR patients.Deep venous catheters (x2 =5.923,P=0.015),invasive mechanical ventilation (x2 =16.845,P=0.000),other catheters (x2 =4.009,P=0.045) and surgery (x2 =3.910,P=0.048) were all significantly different between CRKP and non-CRKP patients.APACHE Ⅱ scores were performed in all patients.The average APACHE Ⅱ score was 8.74-±5.32 of the 50 cases (67.6％) in the survival group and that was 16.46 ± 6.62 of the 24 cases (32.4％) in the death group.The APACHE Ⅱ score in the survival group was significantly lower than that in the death group.The difference was statistically significant (t=5.091,P=0.000).APACHE Ⅱ ≥15 was the independent factor of death (B =-2.708,P=0.000).Conclusions The situation of drug-resistant KP-BSI is severe in the clinic.According to the clinical data,nosocomial infections,invasive mechanical ventilation and deep venous catheters may be the risk factors for MDR bloodstream infection.Deep venous catheters,invasive mechanical ventilation,other catheters and surgery may be the risk factors for bloodstream infection with CRKP.APACHE Ⅱ ≥15 is the independent risk factor for death.The evaluation of APACHE Ⅱ score may predict the prognosis of patients with bloodstream infection.

A new silver amplification immunochromatography system compared with conventional rapid antigen assay for the diagnosis of influenza A and B / 中国感染与化疗杂志

Objective To analyze the consistency of two rapid antigen assays for the diagnosis of influenza A and B. We evaluated the clinical usefulness of silver amplification immunochromatography influenza virus detection kit.Methods Nasopharyngeal swab samples were collected from patients who were suspected of influenza at Huashan Hospital between January and February 2015. Two samples were collected from the same one patient. The samples were tested simultaneously by using IMMUNO AG Cartridge Flu AB kit (AG1) and commercial immunochromatographic assay kit, Clearview exact influenza A&B (CV). PCR method was used as reference.Results A total of 91 samples were tested, of which 7 were positive for influenza A and 53 positive for influenza B by AG1 system; 7 positive for influenza A and 50 positive for influenza B by CV system; and 8 positive for influenza A and 60 positive for influenza B by PCR. The sensitivity and specificity of the AG1 system were 87.5 % and 100 % for influenza A, and 88.3 % and 100 % for influenza B; while the CV system showed sensitivity and specficity of 87.5 % and 100 % for influenza A, 83.3 % and 100 % for influenza B. The AG1 system was 100 % consistent with the CV system in the positive rate of influenza A, and 94.3 % consistent with the CV system in the positive rate of influenza B.Conclusions The AG1 system is well consistent with the conventional immunochromatography-based diagnostic tests in diagnosis of influenza. The AG1 system is useful in earlier diagnosis of influenza due to fewer human error in result interpretation.

Analysis of distribution characteristics of clinical isolates of nontuberculous mycobacteria in a general hospital / 中华传染病杂志

Objective To investigate the current prevalence of nontuberculous mycobacteria (NTM) in Shanghai,and to study the distribution characteristics of NTM clinical isolates,which may help to improve the diagnostic level of NTM and provide guidance for effective prevention and treatment of NTM infection.Methods Culture-positive isolates of clinical mycobacteria were collected from 2008 to 2013 in Huashan Hospital affiliated to Fudan University.All isolates were heat inactivated,and the genomic DNA was extracted and the species were identified by comprehensive comparative analysis of 16S rDNA,hsp65 and rpoB target genes sequencing.Results From January 2008 to December 2013,the overall mycobacterial culture-positive rate was 4.1 ％ (411/10 015).After excluding the repeated isolates,a total of 253 culture-positive mycobacteria isolates were collected for the species identification.By genes sequencing analysis,140 isolates were identified as mycobacterium tuberculosis complex (MTBc),102 NTM and 11 Nocardias,accounting for 55.3％,40.3％ and 4.4％,respectively.Positive rate of NTM isolates had an increasing trend from 25.0％ in 2008 to 42.7％ in 2013,reaching a highest rate of 54.9％ in 2012.In further analysis of 102 NTM isolates,16 species were identified.Among them,28 were M.abscesses,18 strains of M.marinum,17 strains of M.avium-intracellulare complex and 10 strains of M.fortuitum,accounted for 27.5％,17.6％,16.7％ and 9.8％,respectively.Conclusions Both of the isolation number and isolation rate of NTM in the general hospital are increasing.NTM related cases are also increasing in recent years,which mainly caused by M.abscess,M.marinum,M.aviumintracellulare complex and M.fortuitum.

Fatal case caused by H7N9 avian influenza A virus in Shanghai: a case report and literature review / 中华传染病杂志

Objective To understand the epidemiology and clinical features of avian influenza,improve the prophylaxis and treatment.Methods Clinical data of a fatal case caused by H7N9 avian influenza A virus in Shanghai was retrospectively reported and analyzed,literature on avian influenza A virus infection in human was reviewed.Results A severe case of H7N9 avian influenza was reported,with typical clinical characteristics.The epidemiology history of the patient was unclear,all the contacts were tested negative for H7N9 avian influenza A virus.Literature search,H7 subtype of avian influenza in 2012 was only sporadic,the majority of patients with mild symptoms.People did not have immunity against avian influenza.Conclusions Severe case of H7N9 avian influenza progress quickly and its infection pattern is not clear up-to the time point.It needs further exploration and discovery.

Risk factors and prognostic analysis of cranial nerve impairment in 121 patients with tuberculous meningitis / 中华传染病杂志

Objective To investigate the risk factors associated with cranial nerve impairment in patients with tuberculous meningitis.Methods A total of 121 patients with tuberculous meningitis who were admitted to Huashan Hospital from 2000 to 2011 were reviewed retrospectively.Demographic data (gender,age),course of disease,initial results of cerebral spinal fluid (CSF) tests,occurrence of cranial nerve impairment and prognosis of these patients were collected.All the patients were followed up for at least 3 months,and for those with cranial nerve impairment,the minimum follow-up period was 1 year in order to judge the recovery of cranial nerve impairment.Multivariate analysis was performed to study the associated risk factors.Results Out of 121 patients,22 (18.2 ％)developed cranial nerve impairment.Nerves involved were abducens nerve,oculomotor nerve,optic nerve and auditory nerve,and impairment of single nerve occurred in 9 (40.9 ％),8 (36.4 ％),7(31.8％) and 1(4.5％) patient,respectively.Three cases had more than one group of cranial nerves involved,accounting for 13.6％ of the 22 patients with cranial nerve impairment.The incidence of conscious disturbance was significantly higher in patients with cranial nerve impairment than those without impairment (77 ％ vs 45 ％,P=0.020).Delay in diagnosis (OR =1.017,95 ％ CI:1.001-1.033,P=0.040) and occurrence of conscious disturbance (OR =3.242,95 ％ CI:1.142-9.205,P=0.027) were independent predictive factors of cranial nerve injury.During one-year follow-up,90.9％ of patients were fully recovered from cranial nerve impairment,with a median duration of 1 month (range 0.5-6.0 months).Conclusions Cranial nerve impairment is a common complication in patients with tuberculous meningitis.Delay in diagnosis and occurrence of conscious disturbance were independent predictive factors.Most cranial nerve impairment were reversible,and timely diagnosis and treatment are important ways to reduce complications.

The predictive value of drug resistance gene detection for the prognosis of patients with pulmonary tuberculosis / 中华传染病杂志

Objective To identify drug resistance status of Mycobacterium tuberculosis (MTB) strains by the GenoType MTBDRplus line-probe assay (LPA),compare its performance with traditional drug susceptibility testing (DST),and to assess its predictive value for the prognosis of patients with drug resistance tuberculosis.Methods Pulmonary tuberculosis patients who visited Zhuji People's Hospital,Zhejiang Province during February 2011 and January 2012 with a positive result of sputum smear at baseline were all recruited.A total of 275 culture positive specimens were collected,then isolated and cultured for Mycobacterium tuberculosis in the laboratory.DST were performed,meanwhile,GenoType MTBDRplus were also applied to detect resistance to isoniazid (INH) and rifampin (RMP).All the tuberculosis patients who were recruited were followed,including sputum culture and chest radiography.Results There were 192 strains showing drug resistance both by DST and MTBDRplus LPA.Fourteen multidrug resistant (MDR),21 INH mono-resistant and 2 RMP mono-resistant strains were detected by DST.As for GenoType MTBDRplus LPA,MDR,INH mono-resistant and RMP mono-resistant strains were 14,18 and 2,respectively.Taken DST as the gold standard,LPA was more accurate in the detection of resistance to RMP,while it failed to detect 23.8％ (5/21) of the INH-resistant strains.We analyzed the prognosis of patients with drug resistance by GenoType MTBDRplus LPA,the rates of treatment success were 84 ％ (110/131),9/15,3/11 in patients infected with susceptible,INH mono-resistant and MDR strains,respectively.For the 2 cases of RMP mono-resistanee,one was cured and the other failed.The predictive value of molecular drug resistance test for treatment failure in INH mono-resistant patients was 40.0 ％,while that was 83.5 ％ for treatment success in INH susceptible patients.The predictive value for treatment failure in RMP mono-resistant patients was 50.0％,while that was 81.5％ for treatment success in RMP susceptible patients.The predictive value for treatment failure in MDR patients was 72.7％,while that was 81.1％ for treatment success in patients without MDR.Conclusion The GenoType MTBDRplus LPA assay is a rapid and reliable diagnostic test for resistance of MTB,which can be used to predict the prognosis of drug resistant tuberculosis in the clinical practice.

Enzyme-linked immunospot assay for the diagnosis of latent tuberculosis infection in rheumatic diseases / 中华风湿病学杂志

Objective To evaluate a new enzyme-linked immunospot assay (TSPOT-TB) for the diagnosis of latent tuberculosis infection in patients with rheumatic diseases.Methods The rapid TSPOT-TB assay was applied to detect ESAT-6 and CFP-10 specific T cells in blood samples from 126 rheumatic disease patients.A PPD skin test was performed on all patients simultaneously.Results The positive rate of TSPOT assay was 23.8% and that of PPD skin test was 34.9%.The overall agreement between the 2 tests was 71.4%.Among PPD (-) patients (n=82),11 were TSPOT (+) ( 13.4% ).Among PPD (+) patients (n=44),25 were TSPOT(-) ( 56.8% ).The patients who got BCG vaccination showed a significantly higher rate of positive results of PPD skin test than those who did not(41% vs 19%,P＜0.05).While in TSPOT assay,the BCG vaccination did not show any influence on TSPOT results (22% vs 27%,P＞0.05).Conclusion BCG vaccina-tion affects the results of PPD test in patients with rheumatic diseases,but has no influence on TSPOT results.The infection rate of latent tuberculosis in patients with rheumatic diseases in our study is 23.8% detected by TSPOT.

Impaired Vγ2Vδ2+T lymphocytes responses during Mycobacterium tuberculosis infection in human immunodeficiency virus infected individuals / 中华传染病杂志

Objective To evaluate the number and function of peripheral Vγ2Vδ2+T lymphocytes during Mycobacterium tuberculosis(MTB)infection in human immunodeficiency virus (HIV)infected individuals.Methods Seventy-six HIV/acquired immune deficiency syndrome(AIDS) patients co-infected with MTB were divided into active tuberculosis(TB)group(HIV+TB)and latent TB group(HIV+LTB).T cell subsets of peripheral blood lymphocytes were analysed by flow cytometry.Stimulated by protein purified derivative(PPD)and hydroxymethylbutenyl diphosphate (HMBPP),specific interferon(IFN)-γ producing T cells were detected using enzyme-linked immunospot(ELISPOT)and intracellular cytokine staining(ICS).Data were analyzed by t test.Results The absolute number of CD3'T cells and DroDortion of V72VB2'T cells in CD3+T cells in HIV+TB group were both significantly lower than those in HIV+LTB group(t=-3.67,P<0.01;t=-2.06,P<0.05).PPD-specific IFN-y-producing T cells and percentage of PPD-specific CD4+Tcells in CD3+T cells in HIV+LTB group were both similar with those in HIV+TB group.While HMBPP-specific IFN-γ-producing T cells and percentage of HMBPP-specific Vγ2Vδ2+T cells in CD3+Tcells in HIV+LTB group were both higher than those in HIV+TB group(t=2.71 and t=3.003,respectively;both P<0.0 1).Conclusion The number and function of Vγ2Vδ2+T cells were impaired in HIV/AIDS patients coinfected with active MTB infection,which indicates that Vγ2Vδ2+T cells may be the key immune cells against MTB in individuals with impaired CD4+T cells.

Genotypic characteristics of hepatitis B virus isolated from 60 Tibet patients / 中华传染病杂志

Objective To investigate the genotypic characteristics of hepatitis B virus(HBV) in Tibet.Methods Serum samples from 60 cases of hepatitis B in Tibet Autonomous Region were collected from January 2000 to March 2004.HBV genotypes were analyzed by sequencing S region and precore/core region.Results HBV isolated from 60 cases were all found to be D genotype by S region sequencing.Dc mixture was found in 59 cases,showing the recombination between their precore/core gene and genotype B virus at 1804-2299 nucleotide.The other one case showed Dbc mixture genotype,showing recombination between its precore/core gene and genotype B and C genes.Conclusions All Tibet cases in this study show mixture genotype D with recombination with genotvpe C or both genotype B and C at precore/core region.No case of pure genotype D is found.

New multiplex allele-specific PCR assay targeting katG codon 315 variation for detection of isoniazid resistant Mycobacterium tuberculosis strains / 中华检验医学杂志

Objective To develop a new multiplex allele-specific PCR(MAS-PCR) assay to detect mutation in codon 315 of katG gene of Mycobacterium tuberculosis , mutation in this codon has been reported to be able to account for the Mycobacterium tuberculosis clinical isolates resistant to isoniazid(INH).Method Based on the sequence of katG gene of Mycobacterium tuberculosis , three specific primers are designed to carry out the MAS-PCR, 84 purified DNA preparation with known katG 315 variation detected by PCR-restriction fragment length polymorphism (PCR-RFLP) are used to optimize PCR.Results 84 Mycobacterium tuberculosis clinical stains are detected by the MAS-PCR and PCR-RFLP, respectively.The sensitivity of detection by MAS-PCR is 77.8%,and the specificity is 95.2%.katG mutation S315N(AGC→AAC), neglected in RFLP, can be detected by MAS-PCR.Conclusion MAS-PCR assay is sensitive, specific, economic and easy to carry out , can be used in clinical laboratories to detect the INH-resistant Mycobacterium tuberculosis strains.

The investigation of the variability of katG gene in Mycobacterium tuberculosis / 中华传染病杂志

Objective To investigate the variability of katG gene in Mycobacterium tuberculosis strains isolated in China,and screen the new isoniazid resistance related mutation sites meanwhile. Methods 429 clinical Mycobacterium tuberculosis isolates were included in our research.PCR-RFLP method was applied to screen for the S315T mutation firstly.Whole katG gene sequencing was done in those resistant strains without S315T mutation to explore the unknown mutated sites associated with isoniazid resistance.Results S315 mutation were found in 76.9% (166/216)resistant stains. Complete or part deletion of katG gene was detected in 2 highly-resistant isolates.Sequencing in 48 resistant strains showed that 315,463 and 234 sites were the most frequent mutation sites,other sites were also found but distributed dispersedly with low prevalence rate as less than 5%.Besides S315T, S315N were also common in China (8.7%).The most common variation is still site mutation.Con- clusions The results from the study of genotypes associated with most common clinical resistant phe notypes can be helpful to develop new methods to detect the resistant M.tuberculosis.

Simultaneous detection for isoniazid and rifampin-resistanct Mycobacterium tuberculosis by Multiplex Allele-Specific PCR assay / 中华传染病杂志

Objective To develop a new Multiplex Allele-Specific polymerase chain reaction(MAS-PCR) assay to detect the main point mutations in the katG and rpoB genes, which has been reported to account for the majority of clinical Mycobaterium tuberculosis resistant to isoniazid and rifampin. Methods Based on the sequences of katG and rpoB genes, specific primers were designed to carry out the MAS-PCR to detect the most common mutations in codon315 of katG and codons 531,526,516 of rpoB gene. Results The purified DNA preparation of 96 clinical Mycobacterium tuberculosis strains were used to optimize PCR. No mutation was detected in 19 isoniazid-sensitive strains. The 315Ser point mutation was detected in 79.2%(61/77)of isoniazid-resistant strains, the type of mutation includes the most common S315T and the less common S315N, which could’t be detected by PCR-RFLP(polymerase chain reaction-restriction fragment length polymorphism). However, S315G couldn’t be detected by MAS-PCR and that will make a false negative. The mutations in codons 531,526,516 were detected by the MAS-PCR. Compared with the results of direct sequencing of rpoB gene, no mutation was detected in sensitive strains. For rifampin-resistant strains, the total sensitivity was 81.5%(66/81). Conclusions MAS-PCR is a new molecular method with high sensitivity and specificity, which can be used to detect the point mutation in katG and rpoB gene rapidly and economically. It can be used in clinical laboratories to detect drug-resistant tuberculosis strains. Simultaneous detection for katG and rpoB gene mutations in one MAS-PCR system will help to improve the efficiency of this method.