Detection of Aspergillus species in blood and bronchoalveolar lavage samples from immunocompromised patients by means of 2-step polymerase chain reaction: clinical results.

Bronchoalveolar lavage (BAL) samples from 67 patients who were at high risk for invasive aspergillosis were examined using a recently developed 2-step polymerase chain reaction (PCR) that detects 相似文献   

Detection of Borrelia burgdorferi by DNA amplification in synovial tissue samples from patients with lyme arthritis

Objective. To compare the detection rates of chromosomal flagellin gene from Borrelia burgdorfen in synovial tissue (ST) and synovial fluid (SF) using polymerase chain reaction (PCR) techniques. Methods. B burgdorferi DNA was sought in SF and ST from 12 consecutive patients with Lyme arthritis and from 29 patients with noninfectious diseases (controls). Results. No DNA amplification was observed in samples obtained from the 29 control patients, whereas B burgdorferi DNA was detected in all ST and/or SF samples from the 12 patients with Lyme arthritis. Results from 1 ST sample were not interpretable because of PCR inhibitors. Among the 11 remaining patients, 10 had positive ST samples, whereas only 4 had positive SF samples (P < 0.05). Conclusion. These data suggest that detection of chromosomal B burgdorferi DNA may be more efficient in ST than SF.     

Conventional respiratory viruses recovered from immunocompromised patients: clinical considerations.

Lower respiratory tract infection is the most common complication in the immunocompromised patient. From January 1991 to December 1995, 785 consecutive patients with suspected respiratory tract infections were studied. One hundred ninety-nine viruses were isolated from 182 (23%) of 785 bronchoalveolar lavage fluid specimens. Cytomegalovirus was isolated from 131 patients, herpes simplex virus was recovered from 31, and conventional respiratory viruses (CRVs) were recovered from 36. There were 9 influenza A viruses, 2 influenza B viruses, 7 parainfluenza viruses, 5 respiratory syncytial viruses, 5 adenoviruses, 6 enteroviruses, and 3 rhinoviruses. We identified 22 patients from whom a CRV was the only microorganism recovered; 13 patients developed pneumonia, 10 had acute respiratory failure, 5 required support with mechanical ventilation, and 5 (23%) died. In conclusion, CRVs are frequent causes of respiratory illnesses and are associated with high rates of morbidity and mortality among immunocompromised patients.     

Detection of kaposi sarcoma-associated herpesvirus DNA in saliva and buffy-coat samples from children with sickle cell disease in Uganda

Among 233 children, Kaposi sarcoma-associated herpesvirus (KSHV) DNA was detected in 43% of children seropositive for both K8.1 and orf73, in 29% of children seropositive for K8.1 only, in 14% of children seropositive for orf73 only, and in 7% of children seronegative for both K8.1 and orf73; among 228 mothers, KSHV DNA was detected in 27%, 25%, 4%, and 1%, respectively. KSHV DNA was detected more frequently and at higher levels in saliva than in buffy-coat samples and in children than in mothers. In both children and mothers, detection in saliva was associated with detection in peripheral blood. Detection was associated with K8.1 seropositivity, younger age, and high household density, indicating the importance of in-household person-to-person transmission, likely via saliva.     

Detection of bacterial DNA in serial synovial samples obtained during antibiotic treatment from patients with septic arthritis.

OBJECTIVE: The management of septic arthritis could benefit from sensitive tests that detect the persistence of microorganisms in the joint. The aim of this study was to determine the feasibility of monitoring the presence of bacterial DNA in synovial samples from septic arthritis patients during antibiotic treatment. METHODS: Synovial fluid (SF) and synovial tissue (ST) samples were collected serially from 6 patients with septic arthritis before and during antibiotic therapy. In addition, peripheral blood (PB) samples were available for polymerase chain reaction (PCR) analysis from 5 of the 6 patients before treatment. All samples were analyzed for the presence of bacterial DNA with the use of a PCR with universal 16S ribosomal RNA primers. Automated sequencing and comparative data analysis were performed to identify the species. These data were compared with Gram staining and culture results. RESULTS: The bacterial species cultured from the synovium could be identified in all 6 patients using PCR and subsequent sequence analysis of the amplicons. In virtually all cases, positive Gram stain and culture findings in the synovial samples became negative after 2-3 days of antibiotic treatment. Bacterial DNA persisted in the SF and/or ST after culture conversion; in 2 patients, bacterial DNA was still detected at day 10, in 1 patient, at day 20, and in another patient, at day 22 after the initiation of treatment. Synovial samples were available for PCR analysis from 2 patients at day 26. At this time point, bacterial DNA could not be detected anymore. All PB samples were negative by both culture and PCR analysis. CONCLUSION: PCR analysis can be used to monitor the presence of bacterial DNA in synovial samples from patients with septic arthritis during antibiotic treatment. The absence of bacterial DNA could help in the decision to discontinue antibiotic treatment.     

冠心病患者HCMV抗体和HCMV-DNA的测定及其临床意义

目的　探讨人巨细胞病毒 (HCMV)感染与冠心病的关系及 HCMV感染可否作为冠心病的独立危险因子。方法　间接酶联免疫吸附试验 (ELISA)测定 HCMV- Ig G、HCMV- Ig M和 HCMV- Ig A抗体 ,聚合酶链反应 (PCR)测定 HCMV- DNA。结果　冠心病组的 HCMV- Ig G、Ig M、Ig A抗体阳性率、S/N值 (标本 450 nm吸光度值 /阴性对照 450 nm吸光度值 )和 HCMV- DNA阳性率均明显高于对照组 ,且这些指标不受血清胆固醇、甘油三酯水平及是否伴有高血压和 /或糖尿病的影响 (除 HCMV- Ig M抗体阳性率在是否伴有高血压和 /或糖尿病两组间比较 P<0 .0 5外 ,余均为 P>0 .0 5)。结论　 HCMV感染可能与冠心病有关 ,HCMV感染有可能作为冠心病的独立危险因子。     

不同免疫状态下肺曲霉菌感染24例临床分析

目的探讨不同免疫状态下肺曲霉菌感染的临床特点、治疗及预后因素。方法回顾性分析2000年1月至2010年3月北京协和医院有组织病理或者培养证据确诊肺曲霉菌感染的24例病例资料。结果免疫正常组7例,免疫缺陷组17例。和免疫正常组相比,免疫缺陷组病程短(<1个月更多见,88.2%对14.3%),发热症状常见(88.2%对28.6%),咯血少见(11.8%对71.4%),差异均有统计学意义。免疫正常组影像表现为单发病灶者占57.1%高于免疫缺陷组患者(11.8%,P<0.05)。免疫缺陷组主要用药物治疗,免疫正常组中57.1%手术切除。免疫正常组均存活;免疫缺陷组病死率为52.9%,并且死亡可能与存在严重合并症,免疫缺陷宿主因素不能纠正以及外周血淋巴细胞下降等因素有关。结论不同免疫状态的肺曲菌感染患者临床表现及影像表现不同。免疫缺陷的肺曲霉菌感染患者预后差,其中有严重合并症、宿主因素不能纠正、外周血淋巴细胞下降可能与预后不良有关。治疗应根据患者的免疫状态而有所不同。     

免疫功能低下患者肺部并发症的诊治难点与临床策略

近年来我国造血干细胞和实体器官移植取得了突破性进展,移植手术的数量已跻身世界前列;得益于放疗、化疗和综合治疗的进步,肿瘤患者生存期大为延长;风湿病和其他免疫相关疾病的诊治水平提高,患者为维持缓解症状需要长期乃至终生使用糖皮质激素(简称激素)和(或)免疫抑制剂;随着流行病学趋势改变,亚洲获得性免疫缺陷综合征患者数量正处于快速增长期.由于以上种种因素,我国免疫功能低下人群正在不断增长,兼之人口基数大,免疫功能低下患者的绝对数量居世界之首.     

Lung abscess in adults: clinical comparison of immunocompromised to non-immunocompromised patients

Information related to the clinical characteristics and isolated microbes associated with lung abscesses comparing immunocompromised (IC) to non-immunocompromised (non-IC) patients is limited. A retrospective review for 1984-1996 identified 34 consecutive adult cases of lung abscess (representing 0.2% of all cases of pneumonia), including 10 non-IC and 24 IC patients. Comparison of age, gender, tobacco use, pre-existing pulmonary disease or recognized aspiration risk factors were not significantly different between the two groups. Upper lobe involvement accounted for the majority of cases, although multi-lobe involvement was limited to IC patients. There were no differences in the need for surgical intervention, and mortality was very low for both groups. Anaerobes were the most frequent isolates for non-IC patients (30%), whereas aerobes were the most frequent isolate for IC patients (63%). Importantly, certain organisms were exclusively isolated in the IC group and multiple isolates were obtained only from the IC patients.Thus, comparing non-IC to IC patients, clinical characteristics may be similar whereas important differences may exist in the microbiology associated with lung abscess. These findings have important implications for the clinical management of these patient groups, and support a strategy to aggressively identify microbial agents in abscess material.     

Focal hepatic candidiasis: a distinct clinical variant of candidiasis in immunocompromised patients

Focal hepatic candidiasis is a distinct clinical variant of candidiasis in immunocompromised hosts. Although affected patients may exhibit previous evidence of extra-hepatic candidal infection, manifestations of disease at the time hepatic involvement is documented are localized to the liver. Five cases of focal hepatic candidiasis in immunocompromised patients were diagnosed antemortem in the past 15 months at our institution. Clinical, laboratory, and radiologic findings from those five patients, as well as from 20 additional cases reported in the medical literature, are reviewed. Patients with focal hepatic candidiasis had fever unresponsive to antimicrobial therapy, prominent gastrointestinal symptoms and signs, elevated levels of alkaline phosphatase, and hepatic, with or without accompanying splenic, defects noted on abdominal computed tomography. Liver biopsy usually revealed yeast and/or hyphal forms, but cultures were frequently negative. Response to therapy occurred in only 13 (59%) of 22 assessable patients. The apparent increasing incidence of focal hepatic candidiasis may be due to the recent use of more intensely cytotoxic chemotherapies for cancer.     

Correlation between levels of DNA antibodies and clinical disease activity in SLE.

Sera were tested from 23 patients with systemic lupus erythematosus followed over a period of 1 to 5 years. Antibodies to native DNA were measured and correlated retrospectively with clinical evidence of disease activity. The overall degree of correlation between the presence of DNA antibodies and evidence of disease activity was good (P less than 0-001). Of 206 sera tested, only 4 had a normal DNA antibody at a time when significant clinical activity was noted. In contrast, 34 sera had mild to moderately raised DNA antibody levels at times of clinical remission. Although DNA antibodies are a useful investigation in the monitoring of disease activity, changes in therapy should not necessarily be made on DNA antibody levels alone.     

老年慢性阻塞性肺病患者疾病严重程度与血清甲状腺素水平的关系

目的探索老年慢性阻塞性肺病（chronicobstructivepulmonarydisease,COPD）患者血清甲状腺素水平与疾病严重程度的关系。方法用放射免疫法测定了49例健康老年人（对照组）和235例老年COPD患者（包括缓解期和急性加重期患者）血清TT3、TT4、FT4、FT3和TSH水平并对数据进行统计学分析。结果与对照组相比,COPD缓解期患者血清甲状腺激素水平无明显差异,而急性加重期患者甲状腺激素水平明显降低（P〈O．05o急性加重期的三组：无肺心病和呼衰组（A组）、有肺心病无呼衰组（B组）、呼衰组（C组）,B组和C组的甲状腺激素水平基本上都较A组低,但仅有FT3和TT4有显著性差异（P〈0．05）,其中以呼衰组患者最低。另外,FT3水平下降与PaO2下降成正相关。而且COPD死亡组患者除TSH（p〉0．05）外,其余甲状腺激素水平均明显低于存活组（P〈0．05.结论COPD患者存在着明显血清甲状腺激素水平的异常,患者甲状腺激素水平与COPD的严重性、进展和预后有一定关系。     

Correlation between Saccharomyces cerevisiae DNA in intestinal mucosal samples and anti-Saccharomyces cerevisiae antibodies in serum of patients with IBD

AIM: To investigate the correlation between ASCA and presence of mucosal S. cerevisiae DNA in a population of CD, ulcerative colitis (UC) patients and controls. METHODS: S. cerevisiae-specific primers and a fluorescent probe were designed for a 5' exonuclease real time PCR (TaqManTM) assay, which is a homogenous system using a fluorescent-labelled probe for the detection of PCR product in real time. We analyzed the relation of the PCR results with the ASCA findings in a group of 76 inflammatory bowel disease (IBD) patients (31 CD, 45 UC) and 22 healthy controls (HC). RESULTS: ASCA (IgA or IgG) were positive in 19 (61%) patients with CD, 12 (27%) with UC and none of the HC. PCR amplification was inhibited and excluded from the final results in 10 (22%) UC patients, 7 (22%) CD patients, and 6 (30%) HC. In only 15 of the mucosal samples, S. cerevisiae DNA was detected by real time PCR, including 7 (29%) in CD, 7 (19%) in UC, 1 (6%) in HC. In 4 CD and in 4 UC patients, ASCA and mucosal S. cerevisiae were positive. Mucosal S. cerevisiae was present in combination with negative ASCA IgA and IgG in 3 UC, and 3 CD patients. CONCLUSION: We conclude that since the presence of S. cerevisiae in colonic mucosal biopsy specimens is very rare, ASCA is unlikely to be explained by continuous exposure to S. cerevisiae in the mucosa. Therefore, ASCA formation must occur earlier in life and levels remain relatively stable thereafter in immunological susceptible persons.     

Correlation of heart size with clinical and hemodynamic findings in patients with coronary artery disease.

The clinical, hemodynamic, and angiographic findings were correlated with the heart size in 207 patients with proved coronary artery disease. Cardiomegaly was noted in 34 patients and normal heart size in 173. In these two groups, the patients' age range, duration of disease, and history of myocardial infarction were similar. There was no statistical difference in incidence of shortness of breath, hypertension, left ventricular hypertrophy, or abnormal glucose tolerance. Patients with cardiomegaly had a significantly higher incidence of congestive heart failure (26 per cent) as compared to patients with normal heart size (2.9 per cent) (P less than 0.001). Patients with enlarged heart presented a high incidence of anterior wall or multiple myocardial infarction (73 per cent) (P less than 0.001). The cardiomegaly group had a high incidence of elevated end-diastolic volumes, elevated end-diastolic pressures, and diminished ejection fractions when compared to patients with normal heart size (P less than 0.01). Double and triple coronary artery disease was more frequent in patients with cardiomegaly and total coronary score was also higher in this group (P less than 0.005). Asynergy was present in 55 per cent of patients with normal heart size but in 82 per cent of those with enlarged hearts (P less than 0.01). The group of patients with cardiomegaly and documented congestive heart failure had ejection fractions less than 0.30. Cardiac catheterization is probably not advisable in these patients in the absence of associated significant mitral regurgitation, ventricular septal defect, or ventricular aneurysm.     

Laboratory diagnosis of cytomegalovirus infection and disease in immunocompromised patients

PURPOSE OF REVIEW: To review new developments in PCR technology as they apply to detecting cytomegalovirus viremia and pneumonia, recent advances in detecting CMV resistance to antivirals and assays of specific CMV lymphocyte function. RECENT FINDINGS: This review summarizes the attempts to use real time PCR for cytomegalovirus deoxyribonucleic acidemia and to compare it to conventional PCR and antigenemia, it also reviews the use of quantitative PCR on bronchoalveolar lavage to assist in the diagnosis of CMV pneumonia.Phenotypic assays of susceptibility in tissue culture are much too slow to assist clinical decisions, taking weeks for completion. Genotypic assays may be performed directly on clinical samples such as blood, and cerebrospinal fluid and can be done by sequencing in a very few days.Finally, assays of lymphocytic functional responsiveness to cytomegalovirus can be used to identify transplant recipients at continuing risk for cytomegalovirus disease. SUMMARY: Assays for CMV DNA or antigen in blood are superior to culture for documenting viremia and pneumonia. Genotypic assays have largely replaced phenotypic assays for CMV resistance to antivirals. Lymphocyte responses to CMV antigen(s) may identify patients at risk for CMV disease.     

Visceral leishmaniasis in immunocompromised patients with and without AIDS: a comparison of clinical features and prognosis

Visceral leishmaniasis is basically a disease of healthy infants and adults. However, in the last decade an increasing number of cases of kala azar in immunocompromised patients have been reported with emphasis on atypical manifestations of the disease. During a period of 11 years, 20 immunocompromised patients with AIDS (12 patient), haematological neoplasia (3 patients), corticosteroid therapy (3 patients) or renal transplantation (2 patients) were studied by one or more of the authors. We did not find differences in the presentation of leishmaniasis between patient with or without AIDS and most patients had fever, enlargement of the liver and spleen, blood cytopenias and biochemical abnormalities. Serology was more frequently positive in HIV-negative than in HIV-positive patients (100% versus 63.6%; P=0.13). Bone marrow biopsy was diagnostic in 66% and 87% of patients with and without AIDS, respectively. Failure of anti-leishmanial therapy occurred in 6 of 19 patients treated (31.5%), and 3 patients with AIDS and another 3 without AIDS died during the first episode of leishmaniasis. Of 12 survivors, relapses occurred in five (41.6%). Only patients in whom immunosuppression was ameliorated by means of antiretroviral therapy or by reduction of corticosteroid and other immunosuppressive drugs did not relapse. Treatment of kala azar in immunocompromised host is in satisfactory and new drugs or strategies are urgently needed.     

Estimation of salt intake from spot urine samples in patients with chronic kidney disease

ABSTRACT: BACKGROUND: High salt intake in patients with chronic kidney disease (CKD) may cause high blood pressure and increased albuminuria. Although, the estimation of salt intake is essential, there are no easy methods to estimate real salt intake. METHODS: Salt intake was assessed by determining urinary sodium excretion from the collected urine samples. Estimation of salt intake by spot urine was calculated by Tanaka's formula. The correlation between estimated and measured sodium excretion was evaluated by Pearson's correlation coefficients. Performance of equation was estimated by median bias, interquartile range (IQR), proportion of estimates within 30 % deviation of measured sodium excretion (P30) and root mean square error (RMSE).The sensitivity and specificity of estimated against measured sodium excretion were separately assessed by receiver-operating characteristic (ROC) curves. RESULTS: A total of 334 urine samples from 96 patients were examined. Mean age was 58 +/- 16 years, and estimated glomerular filtration rate (eGFR) was 53 +/- 27 mL/min. Among these patients, 35 had CKD stage 1 or 2, 39 had stage 3, and 22 had stage 4 or 5. Estimated sodium excretion significantly correlated with measured sodium excretion (R = 0.52, P < 0.01). There was apparent correlation in patients with eGFR <30 mL/min (R = 0.60, P < 0.01). Moreover, IQR was lower and P30 was higher in patients with eGFR < 30 mL/min. Estimated sodium excretion had high accuracy to predict measured sodium excretion, especially when the cutoff point was >170 mEq/day (AUC 0.835). CONCLUSIONS: The present study demonstrated that spot urine can be used to estimate sodium excretion, especially in patients with low eGFR.     

Reduced natural killer cell activity in patients with systemic sclerosis. Correlation with clinical disease type

Natural killer (NK) cell number and function were determined in 69 systemic sclerosis (SSc) patients (41 with diffuse cutaneous SSc, 24 with limited cutaneous SSc, and 4 with scleroderma in an overlap syndrome). The results were compared with those obtained from 5 patients with Raynaud's disease and from 27 normal controls. Natural and antibody-dependent killing was reduced in the total group of SSc patients compared with controls, but these differences were primarily attributable to patients with the diffuse form of the disease who were seen early in their illness (less than 5 years after onset). NK cell numbers were not significantly reduced in patients compared with controls, although lower numbers were observed in individuals with early diffuse disease. Other clinical parameters, such as treatment with D-penicillamine or the presence of scleroderma-specific autoantibodies, did not exert an independent effect on NK cell function. These findings suggest a possible central role for NK cells in the pathogenesis of SSc.