手部慢性特殊感染病灶中分支杆菌的检测

目的探讨应用Nest-PCR检测技术对普通细菌培养阴性的手部慢性特殊感染患者提供有效的诊治依据。方法对15例手部慢性特殊感染病灶，除一般的病理、普通及特殊细菌培养、直接抗酸染色检查外，应用Nest-PCR技术来检测感染病灶中的分支杆菌，并对分支杆菌进行核酸测序分型。结果15例中普通及特殊细菌培养、直接抗酸染色均呈阴性。病理检查：结核性肉芽肿1例，结核样肉芽肿3例，结节性肉芽肿4例，慢性肉芽肿性炎7例。应用Nest—PCR检测技术，测出9例为非结核分支杆菌(Non-tuberculous myrcobacterium，NTM)阳性，结核杆菌为0。经核酸测序分型属海分支型5例，堪萨斯／胃型2例，鸟分支型1例，龟／脓肿型1例。结论Nest-PCR检测弥补了肺外分支杆菌培养阳性率低下的缺陷，证实了NTM感染，尤其是海分支杆菌远比结核菌常见，是沿海地区手部慢性特殊感染的主要致病因素。     

非结核性分枝杆菌所致手部感染

自从人类发现并使用抗生素以来，引起手部感染的菌群一直在不断发生着一些重要的变化。本文将重点介绍的是近年来不断受到关注的非结核性分枝杆菌所致的手部感染。本病尤其多见于沿海地区经常接触海洋生物或海上作业的人群中，流行病学资料也显示沿海地区分枝杆菌感染率较其它地区高。这里指出的是在沿海地区，尤其是从事渔业的人群当中，除了非结核性分枝杆菌感染较常见，还有许多其它特殊菌群所致手部感染，如非霍乱性弧菌，另外就是伤后继发的各种细菌感染。而且就分枝杆菌的自然分布而言，海水分枝杆菌除了广泛存在于海洋，也同时存在于淡水中。另外几种非结核性分枝杆菌的分布则根本没有沿海与内地的分别。     

沿海地区手部低毒感染病例的临床分析

目的 探讨沿海地区手部低毒性感染的发病机制，治疗和预后。方法 复习24例手部低毒感染病史的发病特点，特征及治疗结果，并分析其病因学及流行病学特点。结果 该病多有海洋生物刺伤史。临床特征与手部分支杆感染的表现相同，但没有结核感染的全身表现。病理学检查多数可见慢性肉芽肿。24例均作病灶清除术，并辅助应用抗分支杆菌的药物化疗。术后用TAM系统评定功能，结果优7例，良9例，可2例，差1例。结论 该病多发于沿海地区，有分支杆菌病的流行因素，病理报告提示与分支杆菌有关。彻底的病灶清除手术可收到较好的疗效。     

Xpert MTB/RIF在骨关节结核患者快速诊断中的应用

目的:探讨Xpert MTB/RIF在骨关节结核患者快速诊断中的应用价值。方法:2014年2月~2014年11月使用Xpert MTB/RIF对49例骨关节结核患者及32例非结核性骨关节病患者的脓液标本进行检测,以临床诊断为金标准,计算Xpert MTB/RIF检测结核分枝杆菌的敏感性、特异性、阳性预测值、阴性预测值、一致率;同时对脓液标本行抗酸染色及结核分枝杆菌快速培养(BACTECT MGIT960),比较Xpert MTB/RIF与抗酸染色、结核分枝杆菌快速培养在敏感性及特异性上的差异;综合上述两方面因素评价Xpert MTB/RIF在骨关节结核患者快速诊断中的应用价值。结果:Xpert MTB/RIF检测单个脓液标本的时间为2.3±0.2h。49例骨关节结核患者脓液标本中,46例Xpert MTB/RIF检测结核分枝杆菌阳性,3例Xpert MTB/RIF阴性;32例非结核性骨关节病患者脓液标本中,1例Xpert MTB/RIF检测结核分枝杆菌阳性,31例Xpert MTB/RIF阴性;以临床诊断为金标准,Xpert MTB/RIF检测结核分枝杆菌的敏感性为93.87%、特异性为96.87%、阳性预测值为97.87%、阴性预测值为91.17%,一致率为95.06%。在46例Xpert MTB/RIF结核分枝杆菌阳性的骨关节结核患者中,10例存在利福平耐药突变基因,耐药突变率为21.73%。49例骨关节结核患者脓液标本中,抗酸染色阳性8例,阴性41例,敏感性为17.39%;结核分枝杆菌快速培养阳性患者11例,阴性38例,敏感性为23.91%;32例非结核性骨关节病患者脓液标本抗酸染色及结核分枝杆菌快速培养均为阴性,其特异性均为100%。Xpert MTB/RIF检测结核分枝杆菌的的敏感性优于抗酸染色及结核分枝杆菌快速培养(P0.05),特异性与抗酸染色及结核分枝杆菌快速培养无明显差异(P0.05)。结论:Xpert MTB/RIF在骨关节结核患者的快速诊断中具有较高的诊断效能,其耗时短,敏感性高,特异性强,与抗酸染色及结核分枝杆菌快速培养比较具有明显优势。     

Xpert MTB/RIF在骨关节结核患者

【摘要】 目的：探讨Xpert MTB/RIF在骨关节结核患者快速诊断中的应用价值。方法：2014年2月~2014年11月使用Xpert MTB/RIF对49例骨关节结核患者及32例非结核性骨关节病患者的脓液标本进行检测,以临床诊断为金标准,计算Xpert MTB/RIF检测结核分枝杆菌的敏感性、特异性、阳性预测值、阴性预测值、一致率;同时对脓液标本行抗酸染色及结核分枝杆菌快速培养（BACTECT MGIT 960）,比较Xpert MTB/RIF与抗酸染色、结核分枝杆菌快速培养在敏感性及特异性上的差异;综合上述两方面因素评价Xpert MTB/RIF在骨关节结核患者快速诊断中的应用价值。结果：Xpert MTB/RIF检测单个脓液标本的时间为2.3±0.2h。49例骨关节结核患者脓液标本中,46例Xpert MTB/RIF检测结核分枝杆菌阳性,3例Xpert MTB/RIF阴性;32例非结核性骨关节病患者脓液标本中,1例Xpert MTB/RIF检测结核分枝杆菌阳性,31例Xpert MTB/RIF阴性;以临床诊断为金标准,Xpert MTB/RIF检测结核分枝杆菌的敏感性为93.87%、特异性为96.87%、阳性预测值为97.87%、阴性预测值为91.17%,一致率为95.06%。在46例Xpert MTB/RIF结核分枝杆菌阳性的骨关节结核患者中,10例存在利福平耐药突变基因,耐药突变率为21.73%。49例骨关节结核患者脓液标本中,抗酸染色阳性8例,阴性41例,敏感性为17.39%;结核分枝杆菌快速培养阳性患者11例,阴性38例,敏感性为23.91%;32例非结核性骨关节病患者脓液标本抗酸染色及结核分枝杆菌快速培养均为阴性,其特异性均为100%。Xpert MTB/RIF检测结核分枝杆菌的的敏感性优于抗酸染色及结核分枝杆菌快速培养（P<0.05）,特异性与抗酸染色及结核分枝杆菌快速培养无明显差异（P>0.05）。结论：Xpert MTB/RIF在骨关节结核患者的快速诊断中具有较高的诊断效能,其耗时短,敏感性高,特异性强,与抗酸染色及结核分枝杆菌快速培养比较具有明显优势。     

253例HIV感染合并慢性腹泻患者粪便标本的病原微生物检测分析

目的 探讨HIV感染者慢性腹泻的病原学种类及其特点,使患者得到及时的预防和治疗.方法 对253例慢性腹泻HIV感染者的粪便标本首先进行了涂片及染色检测,直接涂片检测蓝氏贾第鞭毛虫,采用革兰染色检测真菌孢子及菌丝,抗酸染色(萋-尼氏染色法)检测分枝杆菌,用改良抗酸染色法检测隐孢子虫,采用常规粪便培养检测志贺菌、沙门菌、肠侵袭性大肠埃希菌及真菌,采用微需氧培养检测空肠弯曲菌,采用金标法检测难辨梭状芽孢杆菌毒素(A+B).结果 本研究共检测出各种病原微生物159株,其中真菌最多,占45%(114/253),隐胞子虫占12.6%(32/253),结核分枝杆菌占2.4%(6/253),难辨梭状芽孢杆菌占1.58%(4/253),空肠弯曲菌占0.8%(2/253),沙门菌占0.4%(1/253).结论 真菌、隐胞子虫、分枝杆菌、难辨梭状芽孢杆菌等是HIV感染者慢性腹泻的主要病原菌,积极寻找病原,以期使患者得到及早的预防和治疗.     

非特异性脊柱感染误诊的临床分析

目的：针对误诊为脊柱结核的非特异性感染病例,探讨非结核性脊柱感染的特点,为提高其诊治水平提供临床数据。方法回顾性分析2009年5月～2012年7月收治的23例疑似脊柱结核病例的临床资料。13例患椎部位有明显疼痛;10例有轻度疼痛,活动后加重。影像学检查提示椎体破坏、椎间隙变窄,椎前软组织肿胀。均行病灶清除植骨融合内固定术,并行组织病理学检查、细菌学培养、菌种鉴定、药敏试验等实验室检查,术后应用敏感或广谱抗生素。结果本组随访10～24个月,平均15个月。结核分枝杆菌培养及PCR检测阴性,普通细菌培养阳性10例,其中金黄色葡萄球菌6例、大肠埃希菌2例、克雷伯白菌1例、真菌1例;细菌培养阴性13例。治疗后疼痛均明显好转,神经损害有所恢复。感染得到控制,无复发。结论脊柱非特异性感染的诊断应当结合临床症状,全面有针对性的细菌学检查以及影像学技术或辅以诊断性治疗才能降低其误诊率。     

荧光定量PCR技术检测肺外结核患者样本临床应用

目的评估赛沛分子检测技术（结核分枝杆菌和利福平耐药基因）（GeneXpert MTB/RIF）检测心包和胸腔积液样本中结核分枝杆菌的有效性。 方法收集2015年1月至2017年6月天门市第一人民人民医院和汉川市人民医院收治的286例结核病疑似患者的临床资料,分别收集158例胸腔积液和128例心包积液样品。每个样品均经过抗酸染色涂片镜检、罗氏培养基结核分枝杆菌培养（LJ培养）和GeneXpert MTB/RIF测定。使用结核分枝杆菌罗氏培养作为金标准,评估GeneXpert MTB/RIF技术检测MTB的有效性。 结果在286例积液样本中,MTB通过LJ培养阳性者51例（17.8%）,GeneXpert MTB/RIF测定阳性者43例（15%）,抗酸染色镜检阳性者11例（3.8%）。GeneXpert技术灵敏度、特异性、阳性预测值和阴性预测值分别为84.3%、100%、100%和96.7%,抗酸染色方法的灵敏度、特异性、阳性预测值和阴性预测值分别为18.3%、99.1%、81.8%和85.4%。GeneXpert技术检查心包积液中MTB的灵敏度可达90%。GeneXpert MTB/RIF和抗酸染色镜检鉴定两种样本中结核分枝杆菌有效性的差异具有统计学意义（χ² = 233.199、33.715、P均< 0.001）。 结论GeneXpert MTB/RIF技术对于检测胸腔和心包积液中的MTB具有高灵敏度和高特异性。     

广西HIV感染者呼吸道非结核分枝杆菌定植情况和临床特点分析

目的呼吸道非结核分枝杆菌定植是HIV感染者发生播散性非结核分枝杆菌感染的危险因素,并可能对肺结核的诊断造成干扰。我国HIV感染者中非结核分枝杆菌呼吸道定植情况和临床特点目前尚无较大样本的研究报道。本文分析HIV感染者痰培养中非结核分枝杆菌的阳性率和相关因素,从而总结其临床特点。方法 2006年8月至2008年7月,对广西省4个诊疗机构中CD4〈350的HIV感染者进行临床症状、胸片、痰涂片,痰分枝杆菌培养和血液分枝杆菌培养在内的结核筛查。结果 1073例HIV感染者在结核筛查时进行了痰分枝杆菌培养,检出非结核分枝杆菌87例（8.1%）,129例（12.0%）为结核分枝杆菌。在痰培养阳性的标本中,43%为非结核分枝杆菌。非结核分枝杆菌在呼吸道的定植率随着患者CD4水平降低而逐渐升高,在CD4计数〉200/μl、100～200/μl、50～100/μl和〈50/μl的患者组中分别为2.8%、6.4%、7.1%和9.8%。痰涂片抗酸杆菌阳性的患者中12.0%为非结核分枝杆菌,非结核分枝杆菌呼吸道定植的患者中8.5%痰涂片结果为抗酸杆菌阳性。与痰培养阴性的患者相比,有呼吸道非结核分枝杆菌定植的患者CD4水平较低,除体重下降更多见外,其他临床症状如发热、咳嗽、盗汗、乏力等,以及胸片异常表现均未增加。结论我国HIV感染者中痰培养非结核分枝杆菌的阳性率较高,尤其是在CD4〈200的患者中,需要加强临床观察。在未有进行细菌培养的情况下,非结核分枝杆菌在呼吸道的定植可能造成结核的误诊,应给予重视。     

急性胆源性胰腺炎患者胰液、胆汁中细菌、真菌分布特点及临床意义

目的探讨急性胆源性胰腺炎(ABP)患者胰液、胆汁中细菌、真菌的分布特点及临床意义。方法回顾性分析2019年1月1日至2021年6月30日在宁夏医科大学总医院肝胆外科接受内镜逆行胰胆管造影(ERCP)下胰管支架置入治疗且同期行胆汁、胰液细菌培养的ABP患者的临床资料。共纳入202例患者, 其中男性102例, 女性100例, 年龄(54±16)岁。以是否发生胰腺感染将患者分为两组:胰腺感染组(n=20)和非胰腺感染组(n=182)。在76例胆汁细菌培养阳性患者中, 将60例胰液细菌培养阳性者作为胰液培养阳性组, 16例胰液培养阴性者作为胰液培养阴性组。对比患者培养出的细菌种类及分布、并发症、合并感染情况等临床资料。采用logistic回归分析胰腺感染的相关因素, 绘制受试者工作特征(ROC)曲线分析相关因素的评估价值。结果共送检标本404份(胰液、胆汁各202份), 其中152份(37.6%)培养阳性, 152份阳性标本分离出菌株174株, 革兰阴性菌96(55.2%)株, 革兰阳性菌70(40.2%)株, 真菌8(4.6%)株。与胰液培养阴性组相比, 胰液培养阳性组患者的中性粒与淋巴细胞...     

Nasopharyngeal granulomatous inflammation and tuberculosis complicating undifferentiated carcinoma.

OBJECTIVE: Four cases of nasopharyngeal granulomatous inflammation after radiotherapy for undifferentiated carcinoma were analyzed for tuberculosis, and the histologic features were compared. STUDY DESIGN AND SETTING: We conducted a retrospective study with analysis of tuberculosis by Ziehl Neelsen staining and polymerase chain reaction analysis for Mycobacterium tuberculosis DNA on histologic materials. RESULTS: Three patients had previous nasopharyngeal undifferentiated carcinoma, one had previous metastatic undifferentiated carcinoma to cervical lymph nodes, and all patients received similar radiotherapy regimen. The light microscopic features were similar with epithelioid histiocytes and granulomas with Langhan's giant cells. In 3 cases, acid-fast bacilli were identified by Ziehl Neelsen stain, and 1 was negative. The results of 2 cases were confirmed by polymerase chain reaction analysis for Myocbacterium tuberculosis DNA. CONCLUSION: Granulomatous reaction after radiotherapy of nasopharyngeal undifferentiated carcinoma can be caused by tuberculosis. SIGNIFICANCE: Diligent search for organisms in postirradiation granulomatous inflammation is warranted to avoid missing an occult tuberculosis infection.     

“Triple infections” (fungal,bacterial and viral) in immunosuppressed renal transplant recipients

In a period of 5 years, 8 out of 77 renal transplant patients showed simultaneous fungal, bacterial and viral infections. Candida albicans was found in all cases. The most severe bacterial complications were infections with Klebsiella, Pseudomonas and Staphylococcus aureus. Cytomegalovirus, persistent HBsAg positive hepatitis, herpes zoster, and herpes simplex infections were also found. Seven patients died of bacterial superinfection and miliary tuberculosis. The data presented show that "triple infections" are associated with high mortality and that miliary tuberculosis occurred frequently in immunosuppressed renal transplant recipients.     

The surgical management of atypical mycobacterial soft-tissue infections

Group IV atypical mycobacterial infections, especially Mycobacterium fortuitum and M. chelonei, are being reported with increased frequency. We report our experience with five cases of soft-tissue infection with these acid-fast bacilli. Often these infections are chronic, with formation of abscesses and multiple fistulas. Optimal surgical treatment often requires wide excision of all diseased tissue followed by delayed closure. Presentation, optimal surgical management, and antibiotic therapy are discussed.     

The comparison of pyrosequencing molecular Gram stain, culture, and conventional Gram stain for diagnosing orthopaedic infections.

We have developed a combined real-time PCR and pyrosequencing assay that successfully differentiated the vast majority of gram-positive and gram-negative bacteria when bacterial isolates were tested. The purpose of this study was to evaluate this assay on clinical specimens obtained from orthopedic surgeries, and to prospectively compare the results of "molecular Gram stain" with culture and conventional direct Gram stain. Forty-five surgical specimens were obtained from patients who underwent orthopedic surgery procedures. The DNA was extracted and a set of broad-range PCR primers that targeted a part of the 16S rDNA gene was used for pan-bacterial PCR. The amplicons were submitted for pyrosequencing and the resulting molecular Gram stain characteristics were recorded. Culture and direct Gram staining were performed using standard methods for all cases. Surgical specimens were reviewed histologically for all cases that had a discrepancy between culture and molecular results. There was an 86.7% (39/45) agreement between the traditional and molecular methods. In 12/14 (85.7%) culture-proven cases of bacterial infection, molecular Gram stain characteristics were in agreement with the culture results, while the conventional Gram stain result was in agreement only for five cases (35.7%). In the 31 culture negative cases, 27 cases were also PCR negative, whereas 4 were PCR positive. Three of these were characterized as gram negative and one as gram positive by this molecular method. Molecular determination of the Gram stain characteristics of bacteria that cause orthopedic infections may be achieved, in most instances, by this method. Further studies are necessary to understand the clinical importance of PCR-positive/culture-negative results.     

Antibiotic susceptibility of bacteria infecting total joint arthroplasty sites

BACKGROUND: Currently, there is no consensus regarding the principles of empiric antibiotic treatment of suspected periprosthetic infection following total knee and hip arthroplasties. This study was undertaken to attempt to establish such principles. METHODS: We performed a retrospective analysis of 146 patients who had had a total of 194 positive cultures of specimens obtained at the time of a reoperation following a total knee or total hip arthroplasty at one of two institutions. Patient demographic data, comorbid conditions, bacterial species, the antibiotic sensitivity profile, and the postoperative day on which the culture tested positive were recorded. RESULTS: Specimens from 110 hips and eighty-four knees were positive on culture. Seventy percent of the infections were classified as chronic; 17%, as acute postoperative; and 13%, as acute hematogenous. The mean time between the operation and the positive culture results was three days. Gram-positive organisms caused the majority of the infections. In the series as a whole, 88% of the bacteria were sensitive to gentamicin; 96%, to vancomycin; and 61%, to cefazolin. The most antibiotic-resistant bacterial strains were from patients for whom prior antibiotic treatment had failed. Acute postoperative infections had a greater resistance profile than did chronic or hematogenous infections. Bacteria isolated from patients with a hematogenous infection had a high sensitivity to both cefazolin and gentamicin. CONCLUSIONS: Empiric antibiotic treatment for suspected periprosthetic infection should be guided by the class of the infection and the findings of Gram staining. We believe that, until the final culture results are available, acute hematogenous infections should initially be treated by a combination of cefazolin and gentamicin therapy. All chronic and acute postoperative infections with Gram-positive bacteria and all cases in which a Gram stain fails to identify bacteria should be managed with vancomycin. Infections with Gram-negative bacteria should be managed with a third or fourth-generation cephalosporin. Infections with mixed Gram-positive and Gram-negative bacteria should be managed with a combination of vancomycin and a third or fourth-generation cephalosporin. Furthermore, we believe that if culture results and other confirmatory tests are not positive by the fourth postoperative day, termination of empiric antibiotic therapy should be considered.     

Tuberculosis presenting as carpal tunnel syndrome

A 44-year-old man presented with typical symptoms, signs, and laboratory findings of carpal tunnel syndrome. Mycobacterium tuberculosis was cultured from the flexor tenosynovium excised at surgery. Tuberculosis should be considered in the differential diagnosis of carpal tunnel syndrome with unexplained chronic synovitis. The diagnosis may be missed unless a tissue specimen is analyzed specifically with acid-fast stain and culture. Therapy should include excision of involved synovium, early postoperative mobilization, and appropriate chemotherapy.     

PCR diagnosis on formalin-fixed, paraffin-embedded tissues with acid-fast stain and culture negativity in chronic dialysis patients of cervico-mediastinal tuberculous lymphadenitis

Background: Bacteriologic studies often provide negative results in tuberculous infection, and do not favour early diagnosis. Polymerase chain reaction (PCR) is known to diagnose tuberculosis quickly. With this in mind, we used PCR to detect mycobacterial DNA on formalin-fixed, paraffin-embedded tissues with acid-fast stain and culture negativity in two dialysis patients with cervico-mediastinal lymphadenopathy. Methods: Sections of neck lymph nodes were cut at two different levels. At each level, two semi-adjacent sections with a thickness of 5 &mgr;m each were cut using standard microtomes with disposable blades. The first section mounted on a glass slide was stained b Ziehl-Neelsen, and the second section was examined by PCR based on a 123 bp fragment of IS6110 that is specific for the Mycobacterium tuberculosis complex. Results: The histology of lymph nodes disclosed inflammatory necrotizing granulomas, but acid-fast stain for M. tuberculosis was negative in the two patients. DNA of M. tuberculosis was detected in lymph node samples from each patient by PCR on the IS6110 element and by dot-blot hybridization. Conclusions: PCR assay is a potentially useful approach for early and rapid diagnosis of tuberculous lymphadenitis in chronic dialysis patients, since mycobacterial staining and culture often provide negative results.