Chovanec Zdenek, Veverkova Lenka, Votava Miroslav, Svoboda Jiri, Jedlicka Vaclav, Capov Ivan
1 First Department of Surgery, Medical Faculty, Masaryk University and St. Anne's Faculty Hospital , Brno, Czech Republic .
Surg Infect (Larchmt). 2014 Dec;15(6):786-93. doi: 10.1089/sur.2013.168.
A variety of methods exist to take samples from surgical site infections for cultivation; however, an unambiguous and suitable method has not yet been defined. The aim of our retrospective non-randomized study was to compare two non-invasive techniques of sampling material for microbiologic analysis in surgical practice. We compared bacteria cultured from samples obtained with the use of the swab technique, defined in our study as the gold standard, with the indirect imprint technique.
A cotton-tipped swab (Copan, Brescia, Italy) was used; the imprints were taken using Whatman no. 4 filter paper (Macherey-Nagal, Duren, Germany) cut into 5×5 cm pieces placed on blood agar in a Petri dish. To culture the microorganisms in the microbiology laboratory, we used blood agar, UriSelect 4 medium (Bio-Rad, Marnes-la-Coquette, France), and a medium with sodium chloride (blood agar with salt). After careful debridement, a sample was taken from the incision surface by swab and subsequently the same area of the surface was imprinted onto filter paper. The samples were analyzed in the microbiology laboratory under standard safety precautions. The cultivation results of the two techniques were processed statistically using contingency tables and the McNemar test. Those samples that were simultaneously cultivation-positive by imprint and -negative by swabbing were processed in greater detail.
Over the period between October 2008 and March 2013, 177 samples from 70 patients were analyzed. Sampling was carried out from 42 males and 28 females. One hundred forty-six samples were from incisions after operations (21 samples from six patients after operation on the thoracic cavity, 73 samples from 35 patients after operation on the abdominal cavity combined with the gastrointestinal tract, 52 samples from 19 patients with other surgical site infections not included above) and 31 samples from 11 patients with no post-operative infection. One patient had a sample taken both from a post-operative and a non-post-operative site. Coincidently, the most frequent cultivation finding with both techniques was a sterile one (imprint, 62; swab, 50). The microorganism cultivated most frequently after swabbing was Pseudomonas aeruginosa (22 cases), compared with Escherichia coli when the filter paper (imprint) was used (31 cases). The imprint technique was evaluated as more sensitive compared with swabbing (p=0.0001). The κ statistic used to evaluate the concordance between the two techniques was 0.302. Of the 177 samples there were 53 samples simultaneously sterile using the swab and positive in the imprint. In three samples colony- forming units (CFU) were not counted; 22 samples were within the limit of 0-25×10(1) CFU/cm(2), 20 samples within the limit of 25×10(1)-25×10(2) CFU/cm(2), five within the limit of 25×10(2)-25×10(3) CFU/cm(2), and three of more than 25×10(4) CFU/cm(2).
The hypothesis of swabbing as a more precise technique was not confirmed. In our study the imprint technique was more sensitive than swabbing; the strength of agreement was fair. We obtained information not only on the type of the microorganism cultured, but also on the number of viable colonies, expressed in CFU/cm(2).
存在多种从手术部位感染处采集样本进行培养的方法;然而,尚未确定一种明确且合适的方法。我们这项回顾性非随机研究的目的是比较手术实践中两种用于微生物分析的非侵入性采样技术。我们将使用拭子技术(在我们的研究中定义为金标准)获得的样本培养出的细菌与间接印记技术进行了比较。
使用了一支棉拭子(意大利布雷西亚的科潘公司生产);印记采用沃特曼4号滤纸(德国迪伦的Macherey-Nagal公司生产),剪成5×5厘米的纸片,放置在培养皿中的血琼脂上。为在微生物实验室培养微生物,我们使用了血琼脂、UriSelect 4培养基(法国马恩拉科盖特的伯乐公司生产)以及一种含氯化钠的培养基(含盐血琼脂)。在仔细清创后,用拭子从切口表面取样,随后将同一表面区域印在滤纸上。样本在微生物实验室按照标准安全预防措施进行分析。两种技术的培养结果使用列联表和麦克尼马尔检验进行统计学处理。对那些印记培养呈阳性而拭子培养呈阴性的样本进行了更详细的处理。
在2008年10月至2013年3月期间,对70例患者的177个样本进行了分析。样本取自42名男性和28名女性。146个样本来自手术后的切口(21个样本来自6例胸腔手术后患者,73个样本来自35例腹腔联合胃肠道手术后患者,52个样本来自19例上述未包括的其他手术部位感染患者),31个样本来自11例无术后感染的患者。1例患者的样本取自术后和非术后部位。巧合的是,两种技术最常见的培养结果都是无菌(印记法62例,拭子法50例)。拭子培养后最常培养出的微生物是铜绿假单胞菌(22例),而使用滤纸(印记法)时最常培养出的是大肠杆菌(31例)。与拭子法相比,印记技术被评估为更敏感(p = 0.0001)。用于评估两种技术一致性的κ统计量为0.302。在177个样本中,有53个样本使用拭子法时无菌而印记法呈阳性。3个样本未计数菌落形成单位(CFU);22个样本在0 - 25×10¹CFU/cm²范围内,20个样本在25×10¹ - 25×10²CFU/cm²范围内,5个样本在25×10² - 25×10³CFU/cm²范围内以及3个样本超过25×10⁴CFU/cm²。
拭子法作为更精确技术的假设未得到证实。在我们研究中,印记技术比拭子法更敏感;一致性强度为中等。我们不仅获得了培养出的微生物类型信息,还获得了以CFU/cm²表示的活菌数量信息。