广州市鼠类动物钩端螺旋体的感染调查

目的 调查广州市鼠类动物中钩端螺旋体病的流行情况和遗传特征。方法 笼捕法捕捉鼠类动物,采集脾组织提取总DNA,通过PCR技术检测钩端螺旋体16S rRNA和LipL32基因,测序后进行系统进化分析。结果 采集褐家鼠170只、板齿鼠4只、黄胸鼠3只、黄毛鼠1只和臭鼩鼱22只,共200只鼠类动物,在褐家鼠中检测到钩端螺旋体阳性样本3个,总阳性率1.5%,系统发育分析显示,3份阳性样本序列与致病性问号钩端螺旋体位于进化树上的同一个分支。结论 广州地区鼠类动物中存在致病性问号钩端螺旋体,本地人群存在一定的感染风险。     

钩端螺旋体EMJH培养基实验室应用

目的 探索柯氏培养基和EMJH培养基在培养钩端螺旋体菌的差异及在钩体病防治上应用价值。方法 观察2种培养基在15群15型钩体培养菌培养过程形态、数量、自凝现象等差异,并对健康人临床疑似钩体病人和暴发流行钩体病人血清进行 MAT试验抗体检测。结果 EMJH培养基接种钩体后第2 d就大量生长繁殖,而柯氏培养基1周后才达繁殖高峰,钩体MAT试验也显示EMJH培养基培养的钩体菌敏感性较柯氏培养基培养菌高。结论 EMJH培养基可用于快速分离钩体,有利于早期确定钩体病的流行和实验室研究。     

防污染支气管肺泡灌洗对痰菌阴性肺结核的诊断价值

目的　研究防污染支气管肺泡灌洗(PBAL)技术对痰菌阴性肺结核患者的诊断价值。方法 对96例痰菌阴性肺结核患者进行经支气管镜防污染支气管肺泡灌洗,防污染支气管肺泡灌洗液经过前处理,离心沉渣行涂片荧光染色镜检抗酸杆菌、改良罗氏法分离培养及BACTEC MGIT960快速结核菌培养。结果 防污染支气管肺泡灌洗液离心沉渣镜检抗酸杆菌阳性20例(20.8%),改良罗氏培养基分离培养阳性30例(31.3%),BACTEC MGIT960快速培养技术培养阳性59例(61.5%),最终获病原学确诊62例,确诊率为64.6%。结论 防污染支气管肺泡灌洗对痰菌阴性肺结核患者具有重要的诊断价值,对灌洗液进行多种检查可提高检出的阳性率。     

广州增江沿岸地区鼠形动物及携带病原体调查

目的 了解广州增江沿岸地区鼠形动物及携带伯氏疏螺旋体、钩端螺旋体、恙虫病东方体、埃立克体、新型布尼亚病毒和汉坦病毒6种病原体的情况。方法 使用笼夜法在广州市增城区捕获鼠形动物,利用荧光定量PCR方法检测鼠形动物脏器中的伯氏疏螺旋体、钩端螺旋体、恙虫病东方体、埃立克体、新型布尼亚病毒和汉坦病毒。采用χ²检验和Fisher确切概率法比较不同种属、雌雄和调查点鼠形动物病原体的感染情况。结果 捕获臭鼩鼱和褐家鼠共69只,鼠形动物密度为13.29%。褐家鼠中检出钩端螺旋体为57.89%、恙虫病东方体为31.58%、汉坦病毒为21.05%和伯氏疏螺旋体为5.26%;臭鼩鼱中仅检出伯氏疏螺旋体为2.00%。埃立克体、新型布尼亚病毒均未检出。此外,褐家鼠中存在4种病原体的双重感染,其双重感染率为36.84%。结论 广州增江沿岸地区鼠形动物以臭鼩鼱和褐家鼠为主且密度较高;褐家鼠中病原体感染率高于臭鼩鼱且存在多种病原体的复合感染。当地应加强对鼠形动物尤其是褐家鼠的监测和控制、降低鼠形动物密度。     

江西省致病性钩端螺旋体血清学和分子流行病学研究

目的 对2013年以来江西省致病性钩端螺旋体进行血清学、基因分型分析,以了解江西省钩端螺旋体血清学和分子流行病学特征。方法 对27株钩端螺旋体进行暗视野显微镜凝集试验确定血清群。PCR扩增16S rRNA基因、测序,确定基因种。利用MLST(multilocus sequence typing)研究进行基因分型分析,并应用BioNumerics (Version5.10)软件进行聚类分析。结果 血清群鉴定:27株菌株隶属于4个血清群,其中黄疸出血群为主要优势血清群,占59.26%,其次依次是爪哇群25.92%、澳洲群7.41%和巴达维亚群7.41%。基因种鉴定:27株菌株隶属于L. interrogans和L. borgpetersenii 2个致病性基因种,L. interrogans为江西省主要优势型别,占77.78%。MLST研究显示27株菌株隶属于5个ST型别,其中ST1为主要基因型,占59.26%。BioNumerics软件分析:27株菌株分为5个Clusters对应于5个ST型,MLST基因型别具有明显的地域性特征,而年代间变化不明显。结论 黄疸出血群为江西省主要流行血清群,L. interrogan为主要致病基因种,ST1为主要基因型,充分了解江西省钩体病血清学和分子流行病学特征将对钩体病防控和疫苗制备有一定的指导意义。     

支气管肺泡灌洗液心钠素测定对良恶性肺部疾病鉴别诊断的研究

目的　研究支气管肺泡灌洗液心钠素(ANP)对良性和恶性肺部疾病的鉴别诊断价值。方法 采用放射免疫测定法测定了60例肺良性疾病患者(其中结核30例,炎症30例)和34例肺癌患者支气管肺泡灌洗液心钠素含量。结果 肺癌支气管肺泡灌洗液(BALF)中的ANP水平显著高于肺良性疾病组(P<0.01),有显著性差异。结论 测定BALF中ANP水平是良恶性肺部疾病鉴别诊断的一项有价值的指标。     

海南省不明原因发热病人中钩端螺旋体病的实验检测结果分析

目的了解海南省不明原因发热病人中钩端螺旋体病的感染情况。方法分别用钩端螺旋体显微镜凝集试验(MAT)、酶联免疫试验(ELISA)和聚合酶链反应(PCR)法,检测海南省2015年全省不明原因发热病人的血标本。结果共检测标本179份,其中显微镜凝集试验检出阳性标本40份,感染率为22.35%,男性感染率为21.65%(21/97),女性感染率为23.17%(19/82);菌群中有7个群能检出,分别是问号钩端螺旋体黄疸出血群、拜伦群、秋季热群、波摩那群、流感伤寒群、赛罗群、塔拉索夫群;酶联免疫试验检测阳性38份,阳性率为21.23%,其中IgM阳性6份,检出率为3.35%,IgG阳性32份,检测率为17.88%;血清学检测结果一致。PCR法检测结果均为阴性。结论海南省不明原因发热病人中存在钩端螺旋体的感染,主要型别以黄疸出血群为主。     

不明原因发热与结核病的关系探讨(附136例分析报告)

目的　探讨不明原因发热与结核病的关系。方法 收集1980年1月至2000年7月因不明原因发热住院的258例患者进行回顾性分析。结果 不明原因发热病人中52.7%诊为结核病。结核病与非结核病之间临床表现有显著性差异(P<0.005)。结论 1结核病仍是不明原因发热的主要原因之一。2试验性抗结核治疗不应少于2～2.5个月。     

贵州省钩端螺旋体血清群特异性PCR鉴定结果与分析北大核心CSCD

目的采用血清群特异性PCR技术了解贵州省近年钩端螺旋体(钩体)流行菌群,为贵州省钩体病的防控提供技术手段和科学依据。方法采用致病性钩体特异性PCR方法(G1/G2-PCR)对来自贵州省近年的钩体分离菌株进行鉴定,进一步应用基于致病性钩体O抗原基因的血清群特异性PCR(O-PCR)对贵州省近年的58株钩体分离株进行血清群鉴定,并采用显微凝集试验(MAT)对O-PCR方法的检测结果进行验证。结果 G1/G2-PCR检测结果显示58株菌株均为致病性钩体,O-PCR将58株致病性钩体菌株鉴定为黄疸出血群,与传统的MAT法鉴定结果一致。结论 O-PCR技术可作为我省钩体快速分群鉴定可靠的实验室诊断技术手段,贵州省近年的钩体流行菌群为黄疸出血群。     

贵州省钩端螺旋体血清群特异性PCR鉴定结果与分析

目的 采用血清群特异性PCR技术了解贵州省近年钩端螺旋体(钩体)流行菌群,为贵州省钩体病的防控提供技术手段和科学依据。方法 采用致病性钩体特异性PCR方法(G1/G2-PCR)对来自贵州省近年的钩体分离菌株进行鉴定,进一步应用基于致病性钩体O抗原基因的血清群特异性PCR(O-PCR)对贵州省近年的58株钩体分离株进行血清群鉴定,并采用显微凝集试验(MAT)对O-PCR方法的检测结果进行验证。结果 G1/G2-PCR检测结果显示58株菌株均为致病性钩体,O-PCR将58株致病性钩体菌株鉴定为黄疸出血群,与传统的MAT法鉴定结果一致。结论 O-PCR技术可作为我省钩体快速分群鉴定可靠的实验室诊断技术手段,贵州省近年的钩体流行菌群为黄疸出血群。     

Leptospirosis in Istanbul, Turkey: a wide spectrum in clinical course and complications

Patients with high fever and multiorgan involvement were investigated for the determination of frequency, clinical course and complications of leptospirosis in Istanbul. Leptospirosis was determined in 22 cases among the 35 hospitalized patients that were pre-diagnosed as leptospirosis according to 'Probable Leptospirosis Diagnosis and Follow-up' form. Among the leptospirosis cases 19 were male and 16 were military staff. Mean age was 35.6 y. Dark field examination (DFE), latex agglutination test (LAG), ELISA IgM, leptospirosis culture (LC) and microscopic agglutination test (MAT) were performed to confirm the diagnoses. The most frequent initial symptoms and findings were fever, fatigue, headache, nausea-vomiting and increased muscle sensitivity. Jaundice was noted only in 2 cases. A 74-y-old female patient died after the recurrence of the disease with severe rhabdomyolysis and pulmonary failure. Sagittal sinus thrombosis, perimyocarditis and chronic renal failure were major complications in another 3 patients. ELISA IgM, LC, DFE, LAG and MAT tests were positive in 68, 72, 82, 100 and 100% of the patients, respectively. As a conclusion, diagnosis of leptospirosis is usually overlooked. Clinical awareness, use of probable leptospirosis diagnosis forms and the application of different laboratory methods in the diagnosis of suspected cases may offer the chance to diagnose the leptospirosis accurately.     

Metagenomic next-generation sequencing for the diagnosis of suspected pneumonia in immunocompromised patients

Objectives: To evaluate the potential of metagenomic next-generation sequencing (mNGS), compared with that of comprehensive conventional microbiological tests (CMTs), of bronchoalveolar lavage fluid (BALF) as a front-line diagnostic for immunocompromised patients with suspected pneumonia.Methods: Sixty critically ill immunocompromised patients undergoing both mNGS of BALF and CMTs for suspected pneumonia were retrospectively analysed. The diagnostic performance was compared between mNGS and CMTs, using the composite diagnosis as the reference standard.Results: Forty-nine patients were diagnosed with microbiologically confirmed pneumonia, with 55% having polymicrobial infections. There was no significant difference in the overall diagnostic accuracy between mNGS and CMTs (61.7% vs 76.7%; P = 0.11). mNGS and CMTs had comparable diagnostic accuracy for bacterial and viral infections. Although mNGS identified more viral pneumonia, it had a much lower diagnostic accuracy for fungal infections (76.7% vs 99.2%; P < 0.001), mainly due to the low sensitivity for invasive pulmonary aspergillosis (45.5% vs 100%; P < 0.001).Conclusion: The overall diagnostic performance of BALF mNGS as a first-line diagnostic was similar to that of comprehensive CMTs, except in the case of a lack of consideration of potential pathogens or limited CMTs. The combination of mNGS and CMTs may be the best diagnostic strategy.     

Introduction of a rapid dipstick assay for the detection of Leptospira-specific immunoglobulin m antibodies in the laboratory diagnosis of leptospirosis in a hospital in Makassar, Indonesia

An easy, rapid and robust dipstick assay for detection of leptospira-specific immunoglobulin M (IgM) antibodies was evaluated on 403 patients admitted for hospitalization because of fever. The clinical symptoms and signs of 35 patients were consistent with leptospirosis. The final diagnosis for the remaining patients was as follows: 136 with typhoid fever, 82 with hepatitis, 74 with malaria, 48 with infections of the respiratory tract, and 20 with fever of unknown origin. The clinical diagnosis of leptospirosis was confirmed for 24 (68.6%) patients by the combined results of the microscopic agglutination test (MAT), the reference test for leptospirosis, and of IgM ELISA, a standard laboratory test for the serodiagnosis of leptospirosis. In addition, serum specimens from 8 (2.2%) patients with a final clinical diagnosis other than leptospirosis were found to be positive in MAT and/or IgM ELISA. Compared with the results of MAT and IgM ELISA a sensitivity of 91.6% and specificity of 93.6% was calculated for the dipstick assay. Most of the serum samples from the laboratory confirmed patients gave a moderate to strong staining intensity of the antigen band of the dipstick and were easy to read. The results demonstrate that the dipstick assay is convenient to use and allows the rapid and accurate confirmation of patients with clinical suspicion of leptospirosis in areas where the disease is endemic.     

Real—TimePCR检测棘阿米巴方法建立及应用

目的采用TaqMan探针法建立检测棘阿米巴18srDNA的实时荧光定量PCR（Real—timePCR）方法,为早期诊断棘阿米巴病提供基因诊断标准方法。方法提取实验室培养的4株土壤中分离的和1株水中分离的不同基因型棘阿米巴DNA,测序后在物种保守区域设计Real—time引物和TaqMan探针,用建立的方法检测动物模型兔眼分泌物、大肠杆菌、人巨细胞病毒、卡式肺孢子虫、疑似棘阿米巴角膜炎160例病人眼分泌物。结果与传统实验室培养病原相比,所建立的qPCR检测棘阿米巴角膜炎方法测定大肠杆菌、人巨细胞病毒和卡式肺孢子虫与该研究的实验室分离棘阿米巴物种没有交叉反应,检测结果均为阴性。而160例疑似病人,用培养法检测出感染棘阿米巴患者为5例,用qPCR方法检测出6例,其中5例为培养法测出患者,qPCR方法检测阳性率（3．75％）略高于普通培养法（3．13％）,但无统计学意义（P〉O．05）。qPCR方法在95％置信区间中灵敏度100％（47．95％～100％）高于普通培养法83．33％（36．10％～97．24％）。结论所建立测定棘阿米巴18srDNA的Real-timePCR基因检测方法具有无创伤性、高效、特异、经济等特点,适用于疑似棘阿米巴角膜炎病人门诊筛选的有效方法。     

犬钩端螺旋体病3例报告

目的探究钩端螺旋体病的发病规律和诊疗方法。方法对2015年8月至11月在中国农业大学动物医院确诊的3只钩端螺旋体病犬的发病和诊疗情况进行回顾分析。结果病犬均出现精神沉郁、黄疸、厌食和呕吐等症状,其中2例有体温升高病史,并且在发病前接触过外界水源。实验室检查3只病犬均存在肝肾损伤和贫血。依据尿液PCR检测结果为阳性确诊病犬患钩端螺旋体病。在诊疗过程中3只病犬有1只死亡,其余2只经抗菌素和对症治疗后康复。结论犬感染钩端螺旋体和接触疫水相关,尿液PCR检测可作为该病的确诊方法,治疗上推荐使用青霉素和多西环素等四环素类药物。     

Utility of naproxen in the differential diagnosis of fever of undetermined origin in patients with cancer

The clinical utility of naproxen as an antipyretic agent was examined in the differential diagnosis of fever of undetermined origin in patients with cancer. Twenty-two patients with cancer and fever of undetermined origin for more than seven days were treated with naproxen to control fever when there was no evidence of infection after a careful initial evaluation, and in most cases, after failure of antibiotic therapy. In final analysis, none of five patients with infectious fever had responses to naproxen. In contrast, 14 of 15 patients with neoplastic fever showed a prompt, complete, and sustained lysis of fever within 24 hours after the initiation of naproxen treatment, and the patients also showed symptomatic improvement. One patient with neoplastic fever who did not have a response to naproxen had lysis of fever after the removal of necrotic tumor tissue. Two patients with fever from connective tissue disease had a partial lysis of fever in response to naproxen. These data suggest that naproxen specifically produces the lysis of neoplastic fever and, therefore, is a useful agent in assisting in the differential diagnosis of infectious fever and neoplastic fever in patients with cancer and fever of undetermined origin.     

Factitious and fraudulent fever

The task of elucidating the etiology of fever of undetermined origin remains a major undertaking. Factitious fever is uncommonly considered of major importance in the differential diagnosis of fever of undetermined origin although it is a readily identifiable syndrome and one that is easily excluded once it has been considered. Early identification may reduce the necessity for prolonged, expensive and potentially hazardous hospitalizations for such patients.A retrospective study identified 2.2 per cent (11 of 506) of all patients whose fever on their charts was coded as fever of undetermined origin as having factitious fever. These patients either created factitious fever by manipulation of the thermometer or fraudulent fever by self-induced means. A review of the literature yielded an additional 70 cases in which fever was either the sole factitious sign or part of a larger, more complex factitious illness. Patients were typically young, female and often associated with the medical profession.Patients with factitious fever differ from those with the stereotyped Munchausen's syndrome and may be difficult to recognize. Signs leading to the recognition of this syndrome are emphasized. Since the nature of the psychiatric Illness may vary from patient to patient, early discovery may facilitate psychiatric intervention as such patients may be more amenable to therapy.     

Drug-induced pneumonitis caused by sulfamethoxazole-trimethoprim]

A 33-year-old man was treated with sulfamethoxazole-trimethoprim (SMT-TMP) for an infection in the cervical vertebrae by methicillin-resistant Staphylococcus aureus (MRSA). Two weeks later a fever of 39 degrees C appeared, and a productive cough, hemosputum, and dyspnea developed a further three weeks later. Chest radiographs showed bilateral ground-glass opacity. Cell differentiation of bronchoalveolar lavage fluid (BALF) revealed increases of lymphocytes and eosinophils, and the CD4/CD8 ratio of the BALF lymphocytes was decreased. A thoracoscopic lung biopsy specimen showed fibroedematous thickening of the alveolar walls, hypertrophic alveolar cells, and cell infiltration with neutrophils and lymphocytes. The pathological diagnosis was nonspecific interstitial pneumonia, group II. The fever resolved 6 days after discontinuance of SMX-TMP. The lymphocyte stimulation test for SMX-TMP gave a positive result. Administration of glucocorticoid improved both the symptoms and the laboratory data.     

Application of metagenomic next-generation sequencing for suspected infected pancreatic necrosis

BackgroundMetagenomic next-generation sequencing (mNGS) is increasingly used for the clinical diagnosis of infectious diseases, but there is a paucity of data regarding the application of mNGS in the early diagnosis of infected pancreatic necrosis (IPN).ObjectiveTo investigate the clinical application value of mNGS in the pathogenic diagnosis of IPN.MethodsForty-two patients with suspected IPN were prospectively and consecutively enrolled from August 2019 to August 2021. Blood samples were collected for mNGS and microbial culture simultaneously during fever (T ≥ 38.5 °C). For patients who had indications of surgical interventions, peri-pancreatic specimens were collected for mNGS and microbial culture simultaneously during the first surgical intervention to confirm IPN. The clinical performance of mNGS and microbial culture were compared.ResultsA total of 21 patients (50.0%) were confirmed to have IPN during hospitalization. The sensitivity of blood mNGS was significantly higher than blood culture (95.2% vs. 23.8%, P < 0.001) in diagnosing IPN. The negative predictive value of blood mNGS was 90.0%. The turnaround time of mNGS was significantly shorter than that of microbial culture [(37.70 ± 1.44) vs. (115.23 ± 8.79) h, P < 0.01] and the average costs of mNGS accounted for 1.7% of the average total cost of hospitalization. The survival analysis demonstrates that the positive blood mNGS result was not associated with increased mortality (P = 0.119).ConclusionsWith more valuable diagnostic performance and shorter turnaround time, clinical mNGS represents a potential step forward in the early diagnosis of IPN.     

宏基因二代测序技术对非结核分枝杆菌肺病的诊断价值

目的: 探讨宏基因二代测序技术(metagenomic next-generation sequencing,mNGS)在非结核分枝杆菌肺病(nontuberculous mycobacterial pulmonary disease,NTM-PD)中的诊断价值。方法: 回顾性搜集2020年1月至2022年2月浙江大学医学院附属杭州市胸科医院结核病诊疗中心收治的123例疑似NTM-PD患者的资料。根据诊断标准,123例疑似NTM-PD患者最终诊断为NTM-PD患者(NTM-PD组;74例)和非NTM-PD患者(非NTM-PD组;49例)。123例患者肺泡灌洗液、痰液、肺组织同时进行mNGS、PCR荧光探针法、BACTEC MGIT 960分枝杆菌全自动快速液体培养(简称“液体培养”),比较3种方法的诊断效能。结果: 以最终临床诊断结果为标准,mNGS、液体培养及PCR荧光探针法检测NTM-PD的敏感度分别为83.8%(62/74)、78.4%(58/74)、67.6%(50/74),特异度分别为65.3%(32/49)、91.8%(45/49)、87.8%(43/49),诊断一致率分别为76.4%(94/123)、83.7%(103/123)、75.6%(93/123),Kappa值分别为0.524、0.647、0.521。以液体培养结果为标准,mNGS和PCR荧光探针法检测NTM-PD的敏感度分别为87.1%(54/62)和75.8%(47/62),特异度分别为59.0%(36/61)和85.2%(52/61),诊断一致率分别为73.2%(90/123)和80.5%(99/123),Kappa值分别为0.462和0.587。对74例NTM-PD患者菌株进行mNGS及基因芯片法菌种鉴定,以基因芯片法为菌种鉴定标准,mNGS检测的准确率为63.8%(37/58)。结论: 在NTM-PD的诊断中,mNGS检测的敏感度较高,可直接鉴定至菌种水平,但特异度较低,应重点提高检测特异度,以更好地服务于临床工作。