Biochemical identification and molecular characterization (PCR-RFLP) of Nocardia isolates from sputum

BACKGROUND: Nocardia identification has been based on biochemical and morphological characteristics. However, molecular biology techniques allow a better characterization of species and biotypes that are related to invasive diseases. METHODS: Twelve isolates of Nocardia spp. were obtained from sputum of patients with tuberculosis under retreatment. Identification was done based on morphological characteristics, biochemical tests (casein, tyrosine, xanthine, gelatin, and urea) and molecular biology techniques (PCR-RFLP) using restriction enzymes MspI, HinfI, BsaHI, HaeIII and BstEII. RESULTS: Biochemical tests identified the 12 isolates as Nocardia asteroides. PCR-RFLP technique identified nine isolates to species and biotype level: five as N. asteroides type II, two as N. asteroides type VI, and two as N. asteroides type I. The remaining three isolates were identified as follows: one to species level as N. farcinica and two at genus level as Nocardia sp. CONCLUSIONS: Significant statistical differences between the use of traditional techniques and PCR-RFLP were not found at genus level, but there were important differences at species and biotype level. Biochemical tests identified correctly the actinomycete isolates as belonging to Nocardia genus, but at N. asteroides complex level were not able to discern among their different species. PCR-RFLP is a rapid, non-expensive, and reliable method that allows to discriminate the N. asteroides complex species, identifying biotypes related to invasive disease. Our results suggest that the hospital environment was not a contamination source.     

Infection with Nocardia species in Queensland. A review of 102 clinical isolates.

OBJECTIVE: To review the species distribution, pathologic significance and disease associations of clinical isolates of Nocardia and related bacteria in Queensland, and to examine the characteristics, treatment and outcome of patients infected with these organisms. DESIGN AND SETTING: A retrospective review of Queensland State Health Laboratory records provided microbiological data for Nocardia isolates referred from other laboratories during the period January 1983 to December 1988. Clinical information was extracted from hospital case notes, or obtained from detailed questionnaires completed by attending physicians. Nocardia isolates were classified as "significant" if specific treatment for nocardiosis was given, or on the basis of autopsy findings. PATIENTS: One hundred and two patients had a Nocardia species or a related organism isolated from clinical specimens during the study period. RESULTS: The 102 isolates included Nocardia asteroides (45), N, brasiliensis (35), N. caviae (5) and N. transvalensis (5). Clinical results were available for 93 patients, of whom 74 (80%) had a significant isolate recovered. Primary pulmonary or disseminated disease occurred in 35 patients, and was caused mainly by N. asteroides. Significant infections of skin and soft tissues, primarily due to N. brasiliensis, were found in 39 patients. Preexisting lung disease and treatment with steroids and immunosuppression were risk factors for pulmonary and disseminated nocardiosis. A history of inoculation in an outdoor setting was frequent in patients with cutaneous disease. Antibiotic regimens that included trimethoprim-sulfamethoxazole or another sulfonamide agent were used to treat the majority of patients with significant infection. Deaths were confined to those with pulmonary and disseminated disease, with a case fatality rate of 40% in that group. CONCLUSION: Infection with Nocardia species appears to be more common than is generally appreciated. The local species distribution and disease spectrum are similar to those described elsewhere. A high index of suspicion for nocardiosis should be maintained in susceptible hosts with pulmonary infiltrates, particularly when there is evidence for metastatic infection, and in patients with superficial infections and a history of outdoor injury.     

Primary cutaneous nocardiosis.

Seven patients with primary cutaneous nocardiosis were seen during a 20-month period. Six patients had a history of local trauma to an extremity or did frequent yard work. The spectrum of clinical disease included cellulitis, pustules,pyoderma, or lymphocutaneous syndrome mimicking sporotrichosis. Evidence of Nocardia infection elsewhere was absent, and no patient had a serious underlying disease. Nocardia brasiliensis was isolated from four cases, and N asteroides was identified in the other three patients. In two patients, disease resolved without chemotherapy, while a short course of a sulfonamide with or without trimethoprim therapy was successful in the remaining cases. The relatively benign nature of this disease, the usual failure to obtain cultures of superficial skin infections, and our accumulation of seven cases in less than two years suggest this disease is more common than previously recognized.     

Nocardia asteroides sinusitis. Presentation as a trimethoprim-sulfamethoxazole responsive fever of unknown origin.

A patient with a yearlong fever of unknown origin responded to trimethoprim-sulfamethoxazole and was discovered to have culture-proved Nocardia asteroides sinusitis, with absence of detectable disease in other organs. An inhalational route is postulated as the mode of entry of the organism, with localization in the maxillary sinus.     

Mandibular osteomyelitis and multiple skeletal complications in Albers-Schönberg disease

Albers-Sch?nberg disease, or autosomal dominant osteopetrosis type II, is the most common form of the rare disease, osteopetrosis. Mandibular osteomyelitis is a rare complication of the disease. A host of other skeletal complications may also occur. Mandibular osteomyelitis along with bilateral severe coxa vara and pars fracture is very rare in Albers-Sch?nberg disease. We present the occurrence of these complications in a 24-year-old man with Albers-Sch?nberg disease. His mandibular osteomyelitis was successfully treated.     

MRI在骨髓炎特征性表现诊断中的应用进展

骨髓炎是由细菌感染引起的一种进行性骨骼炎症,是造成全世界儿童和成人永久性残疾的重要原因,对该疾病不同时期的有效诊断是目前临床具有挑战性的问题。影像学在疾病的辅助诊断和指导临床治疗方面发挥着核心作用,特别强调MRI,它能很好地显示解剖细节,有较好的软组织分辨率,并能显示骨外疾病的进展。准确诊断各分期骨髓炎需要对该疾病的关键影像表现有较好的了解。现综述脂肪球征、半暗带征、Brodie脓肿以及死骨相关的影像学特征性表现的应用进展,有助于更好地诊断骨髓炎,提高该疾病诊断的准确率。     

慢性多灶性骨髓炎1例及文献回顾

分析1例慢性多灶性骨髓炎病例的临床表现、血清学及影像学检查、诊断依据、治疗方案、出院后随访评估等,了解慢性复发性多灶性骨髓炎的发病机制、诊断、鉴别诊断、治疗,并进一步了解自身炎症性骨病的特点。本例患者青少年起病,伴有严重皮损,有进行性加重的脊柱关节疼痛,结合临床表现及辅助检查,符合慢性多灶性骨髓炎的诊断。经过抗炎、止痛效果不佳,后换用肿瘤坏死因子α(tumor necrosis factor α,TNF-α)抑制剂后疼痛缓解,炎症指标恢复正常,皮疹及影像学检查明显改善。慢性复发性多灶性骨髓炎属于自身炎症性骨病中多基因疾病的一种,又称慢性非细菌性骨髓炎,是一种罕见的非感染性炎性疾病,可引起多灶性溶骨性病变,以周期性加重和缓解为特征。该病发病率低,发病机制不清楚,可能与促炎及抗炎失衡有关,诊断无特异性指标及统一的诊断标准,常易与代谢性骨病、感染、肿瘤等疾病相混淆,临床表现为骨痛、发热、皮疹、骨折等,实验室检查可见炎症指标明显增高,影像学检查有溶骨性或硬化性改变,MRI在识别骨病变和组织水肿上更有效,而且比骨发射型计算机断层扫描(emission computed tomography,ECT)更准确。治疗用药方面,多数开始使用非甾体类抗炎药,但易复发并出现新发病灶,还可选择其他治疗方案,如糖皮质激素、TNF-α抑制剂及双磷酸盐、甲氨蝶呤等改善病情的抗风湿药物(disease-modifying anti-rheumatic drugs,DMARDs)。对慢性多灶性骨髓炎的早期诊断和治疗可以预防和减少疾病的并发症,改善患者预后。     

Salmonella Osteomyelitis

Salmonella infection can cause four predominant clinical syndromes: enteric fever, acute gastroenteritis, bacteraemia with or without metastatic infection, and the asymptomatic carrier state. Salmonella as an aetiological agent in osteomyelitis is essentially rare and salmonella osteomyelitis in itself is predominantly seen in patients with haemoglobinopathies such as sickle cell disease or thalassemia. There are very few cases reported in the literature in which salmonella osteomyelitis is seen in otherwise healthy individuals. We describe here a case of salmonella osteomyelitis in a young gentleman with no significant comorbidities who presented with fever and severe back pain, having returned from recent foreign travel. It is therefore important to consider uncommon pathogens in the differential diagnosis of travellers with prolonged fever and insidious symptoms.     

Minocycline treatment of pulmonary nocardiosis

Minocycline hydrochloride was used to treat pulmonary infections with Nocardia asteroides in five cardiac allograft recipients. In three patients, minocycline was successfully used as the only antinocardial agent. Two other patients were found to have leukopenia after initial therapy with sulfisoxazole. These two patients were subsequently treated with minocycline. The clinical success with minocycline in these highly immunosuppressed patients suggests that minocycline is an effective antinocardial agent. These data did not allow any conclusion regarding which drug, minocycline or sulfisoxazole, is superior in the treatment of this disease.     

Osteomyelitis following Bacille Calmette-Guerin vaccination

Secondary complications after Bacille Calmette-Guerin (BCG) vaccination are unusual. We describe 3 immunocompetent children who developed osteomyelitis after BCG vaccination. The course of the disease is not dramatic, but marked changes are frequently visible in plain radiograph and MRI. The real-time polymerase chain reaction has an essential role to confirm the diagnosis of BCG osteomyelitis. With proper surgical intervention and chemotherapy, the prognosis is usually good.     

一期手术治疗63例Cierny-Mader Ⅱ型及Ⅲ型慢性跟骨骨髓炎的单中心临床诊疗分析

  目的  慢性跟骨骨髓炎致病因素多,分型复杂,治疗具有挑战性。本研究回顾性分析Cierny-Mader Ⅱ型及Ⅲ型慢性跟骨骨髓炎的临床特点,并探究采用一期手术的临床疗效和影响肢体功能的相关因素。  方法  回顾性分析东部战区总医院2005年1月—2018年12月收治的63例Cierny-Mader Ⅱ型及Ⅲ型慢性跟骨骨髓炎患者,其中男性55例,女性8例; 年龄6~68岁,中位年龄为39(23,57)岁。Cierny-Mader分型:Ⅱ型32例,Ⅲ型31例。32例Ⅱ型患者单纯采取一期病灶清除,18例Ⅲ型患者采取病灶清除后植入自体骨,13例Ⅲ型患者采取病灶清除后抗生素骨水泥填塞。采用AOFAS-AH评分量表对患肢术后功能评估。  结果  63例患者均获随访,随访时间12~68个月。8例患者复发,其中32例Ⅱ型患者中5例复发; 采自体髂骨植入的18例Ⅲ型患者中2例复发; 采取抗生素骨水泥填塞的13例Ⅲ型患者中1例复发。63例患者术前AOFAS评分为(63.4±7.6)分,术后AOFAS评分为(84.8±7.4)分。统计分析后发现术前肢体功能与Cierny-Mader分型(P < 0.001)及跟骨骨髓炎的病因(P=0.005)有关,术后AOFAS评分与患者年龄(P=0.018)及病因(P=0.035)相关。  结论  一期手术治疗周期短,疗效良好,是治疗Cierny-Mader Ⅱ型及Ⅲ型慢性跟骨骨髓炎的有效手段。      

侵袭性假丝酵母菌腰椎骨髓炎一例

假丝酵母菌性椎体骨髓炎是一类相对少见但具有挑战性的临床疾病,其临床表现缺乏特异性,常导致诊断延误,引发严重的并发症,故早期诊断是提高治愈率的关键。本文报道1例胃肠道手术后侵袭性假丝酵母菌腰椎骨髓炎病例,并结合临床特点进行分析、总结,以提高对该疾病的早期诊断与治疗水平。     

骨髓炎动物模型的研究进展

骨髓炎是由于感染引起的骨及其相关组织的急性或慢性炎症。该病具有治疗困难、花费高、病程迁延不愈、致残率高等特点。由于骨髓炎发病过程非常复杂、变异度高,目前有多种动物模型应用于骨髓炎的发病机制、诊断和治疗的研究。建立骨髓炎模型的动物常使用兔、大鼠和小鼠,较少使用狗、猪、山羊等大型动物。然而,目前研究中尚未发现可以较好控制的动物模型。本研究通过检索目前骨髓炎动物模型的建立方法,以期寻求一种可以较好控制的用于研究骨髓炎发生、发展的动物模型,并对目前骨髓炎研究进展进行展望。     

慢性创伤性骨髓炎综合治疗进展

骨髓炎的治疗一直是骨科医师面临的最困难的难题之一。近年来慢性创伤性骨髓炎的发病率显著增加,虽然抗生素的应用和积极的手术治疗,使得病死率及致残率已经明显降低,但是有效的治疗仍相当的困难。中西医对此病的治疗各具优势,该文综合分析近几年慢性创伤性骨髓炎的治疗进展,探讨中西医结合、内外兼顾的综合治疗方法。     

The management of childhood osteomyelitis.

The management of seventy-six children admitted to Aler Hey Children''s Hospital with osteomyelitis is discussed and the indications for surgical intervention are defined. The clinical and pathological course of the disease is examined and the term post-acute osteomyelitis is introduced. The importance of recognition of this stage of the disease and its relevance to the management is highlighted. The use of single antibiotic therapy (clindamycin) is shown to be at least as effective as combinational therapy. Complete resolution was achieved in 94.7% of children, surgical intervention was performed on 74.6%.     

Pott's disease in heroin addicts.

Five cases of tuberculous vertebral osteomyelitis occurred in heroin addicts at St Luke's Hospital Center between 1955 and 1975. All patients were men, and four were between 19 and 33 years of age. In contrast with the indolent onset typical to pyogenic or fungal vertebral osteomyelitis in heroin addicts, patients with Pott's disease had an acute toxic reaction, with fever, back pain, weight loss, and night sweats. Neurologic abnormalities, uncommon with pyogenic or fungal vertebral infection, occurred in all patients with tuberculous involvement of the spine. Roentgenograms of the spine were atypical in four patients, including two with an "ivory vertebra." All patients had involvement of extravertebral sites, and one patient had infection with Mycobacterium tuberculosis and M avium.     

Acute haematogenous anaerobic osteomyelitis in sickle cell disease. A case report and review of the literature

Patients with sickle cell disease are more susceptible to acute anaerobic osteomyelitis due to focal gut mucosal ischaemia, translocation of bacteria, and seeding in infarcted bone marrow. Modulation of the immune system is also present. The isolation of anaerobic organisms requires a high index of suspicion, correct specimen collection procedures and meticulous specimen handling. Bacteroides is the predominant organism isolated. Intra-osseous gas in the bone may be seen within four days and radiographs are therefore useful earlier than with aerobic osteomyelitis. Surgical débridement and intravenous antibiotics are the mainstay of treatment with the erythrocyte sedimentation rate being relied on heavily to guide conversion to oral antibiotics. Coexistence of septic arthritis is more common with anaerobic osteomyelitis.     

持续负压引流治疗慢性骨髓炎23例疗效观察

目的 探讨持续负压引流（VSD）对慢性骨髓炎的治疗效果.方法 收治的慢性骨髓炎合并有慢性溃疡的患者共23例,均采用溃疡清创后,使用VSD治疗,待创面清洁、肉芽组织生长良好后,12例直接缝合,8例植皮封闭创面,3例经邻位皮瓣转移封闭创面.结果 VSD平均吸引15 d,平均更换VSD 2.5次.创面渗液均明显减少,且创面面积均有不同程度的缩小.经二期手术后治愈,随访6~29个月,无复发.结论 VSD治疗对慢性骨髓炎具有引流充分、缩小创面、促进肉芽组织生长、降低复发率等优点,疗效确切.     

An unusual complication of sternal and clavicle osteomyelitis in a child with sickle cell disease

We report an unusual child with sickle cell disease, in which osteomyelitis of the sternum and clavicle was diagnosed at the same time. The standard x-ray failed to demonstrate the lesion. Magnetic resonance imaging was very helpful in locating the site and degree of involvement. We recommend the use of magnetic resonance imaging to delineate such findings.     

Destructive bone disease in early syphilis.

Although destructive bone disease is a well-known complication of tertiary syphilis, osteitis or osteomyelitis are not commonly recognized as complications of early (primary or secondary) syphillis. A patient with secondary syphilis characterized by generalized lymphadenopathy, perianal condyloma lata, and positive rapid plasma reagin (RPR) and fluorescent treponemal antibody-absorption (FTA-ABS) tests also complained of headache, right should pain, and right anterior chest pain and swelling. Roentgenograms showed mottled osteolytic lesions consistent with previously described luetic bone disease. Biopsy confirmed the diagnosis of syphilitic osteomyelitis, and treatment with penicillin resulted in prompt resolution of symptoms.