High prevalence of Chlamydia trachomatis,Neisseria gonorrhoeae and particularly Trichomonas vaginalis diagnosed using US FDA‐approved Aptima molecular tests and evaluation of conventional routine diagnostic tests in Ternopil,Ukraine

Sexually transmitted infections (STIs) remain major public health problems globally. Appropriate laboratory diagnosis of STIs is rare in Ukraine. We investigated the prevalence of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) and Trichomonas vaginalis (TV) using the US FDA‐approved Aptima Combo 2 and Aptima TV assays and compared the results with the conventional routine diagnostic tests (CDTs) in Ukraine. Urogenital swabs from consecutive mostly symptomatic females (n = 296) and males (n = 159) were examined. The prevalences were as follows: 10% (n = 47) of TV, 5.3% (n = 24) of CT and 1.5% (n = 7) of NG. The specificity of some CDTs was high, for example, 100% for NG culture, TV IgG ELISA, CT IgM ELISA and CT microscopy, but lower for other CDTs, that is, from 44% to 99.8%. The sensitivity of all CDTs was suboptimal, that is, 71% (n = 5) for NG microscopy, 57% (n = 4) for NG culture, 53% (n = 8) for CT IgG ELISA, 33% (n = 1) for TV IgG ELISA, 28% (n = 13) for TV microscopy, 25% (n = 1) for CT IgA ELISA, 20% (n = 3) for CT IgM ELISA and 0% (n = 0) for CT microscopy. The prevalences of particularly TV and CT were high, but substantial also for NG, in Ternopil, Ukraine. The sensitivities of all CDTs were low, and widespread implementation of validated, quality‐assured and cost‐effective molecular diagnostic STI tests in Ukraine is imperative.     

2010~2012年913株肺炎克雷伯菌感染的分布与耐药性研究

目的调查肺炎克雷伯菌的分布及其耐药性变化。方法选择2010年1月至2012年12北京市肿瘤防治研究所收集的913株肺炎克雷伯菌临床分离菌株作为研究对象,采用K-B法进行药敏试验,所得数据采用WHONET 5.6进行分析。结果肺炎克雷伯菌临床分离菌株的标本主要来源于痰标本(46.0%);超广谱β-内酰胺酶(ESBLs)阳性菌株的阳性率为51.2%;连续3年的药敏结果显示,肺炎克雷伯菌对哌拉西林、头孢呋辛、头孢噻肟、氨苄西林/舒巴坦、复方磺胺甲口恶唑和环丙沙星有较高的耐药率(>30%)。结论近3年来,肺炎克雷伯菌所致感染的耐药率趋于平稳,但肺炎克雷伯菌ESBLs阳性率较高。     

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??Objective To compare pbp2b??ply and lytA genes PCR with cerebral spinal fluid culture in diagnosis of Streptococcus pneumoniae meningitis and prediction of its susceptibility. Methods A nested PCR targeting pbp2b and another two S.pneumoniae specific PCR targeting the genes of pneumolysin??ply?? and autolysin??lytA?? were developed for detection of S.pneumoniae in cerebral spinal fluid from bacterial meningitis patients. The three PCR results and culture were compared. The consistency of penicillin susceptibility PCR ??using resistant and susceptible primers respectively????sequencing and culture-based phenotypic penicillin resistant results were compared. Results Of the 161 specimens studied??there were 25 cases of S.pneumoniae infection confirmed by different methods??16 by pbp2b PCR??16 by lytA PCR??14 by ply PCR and 9 by cerebrospinal fluid culture??. Of the 16 pbp2b positive specimens??penicillin sensitive and resistant sequence types accounted for half??respectively. Four of the 16 pbp2b positive specimens had culture-based phenotypic penicillin-resistant result. Three of 4 were consistent with penicillin susceptibility PCR result. The results of susceptibility PCR targeting pbp2b was consistent with sequencing result. New sequence types were found but there were no new point mutations in these strains when compared with GenBank ??http??//blast.ncbi.nlm.nih.gov/Blast.cgi??. Penicillin resistance in pneumococcal meningitis was 66.67%??6/9?? by culture phenotype and 50%??8/16?? by PCR and sequencing when culture was negative. Conclusion Pbp2b can serve as a good target gene to detect S.pneumoniae and predict its penicillin susceptibility??which is especially important when culture is negative.     

肺炎支原体感染后小儿血常规变化及临床意义分析

目的观察肺炎支原体感染后小儿血常规的变化,并分析其临床意义。方法筛选出2018年10月-2019年9月于在我院进行诊治处理的60例肺炎支原体感染患儿作为研究对象,并将其作为观察组,同时选择同期到我院进行健康体检的60例患儿作为对照组。记录、对比两组的白细胞计数、中性粒细胞比例、淋巴细胞比例、淋巴细胞绝对值、单核细胞比例、单核细胞绝对值、红细胞计数、血红蛋白、血小板计数等血常规指标水平。结果对检测结果展开分析后可知,两组的红细胞计数、血红蛋白、血小板计数等血常规指标水平对照,无明显差异(P>0.05);相较于对照组,观察组患儿的白细胞计数、单核细胞绝对值、中性粒细胞及单核细胞等血常规指标水平呈现明显的提升趋势(P<0.05),而淋巴细胞比例、淋巴细胞绝对值等血常规指标水平呈现明显的下降趋势(P<0.05)。结论患儿肺炎支原体感染后血常规指标会发生明显变化,对血常规指标进行监测可为临床诊断及治疗提供重要依据。     

纤维支气管镜联合阿奇霉素治疗儿童难治性肺炎支原体肺炎的效果

目的研究纤维支气管镜联合阿奇霉素对儿童难治性肺炎支原体肺炎的疗效。方法将101例难治性肺炎支原体肺炎患儿随机分为对照组(51例,单用阿奇霉素)和观察组(50例,纤维支气管镜联合阿奇霉素)。比较两组的治疗效果。结果观察组的治疗总有效率高于对照组,胸片恢复正常时间、咳嗽恢复时间、C反应蛋白恢复时间及体温恢复时间均短于对照组(P<0.05)。治疗后,观察组的FEV1/FVC和MVV高于对照组,血清IL-6、内毒素、TNF-α和LPO水平低于对照组(P<0.05)。结论纤维支气管镜联合阿奇霉素能有效降低难治性肺炎支原体肺炎患儿的炎症因子水平,改善其肺功能,值得临床推广应用。     

肺炎克雷伯菌对β-内酰胺类抗生素耐药性主决定因素的研究

目的了解肺炎克雷伯菌对β-内酰胺类抗生素耐药性及耐药性特点,并探究其主决定因素。方法对临床送检的痰液、咽拭子、气管分泌物等做细菌培养,根据药敏试验结果分为敏感菌株、泛耐药菌株、多重耐药菌株3种菌株。分析菌株β-内酰胺酶活性以及细胞外膜通透性的变化。结果与敏感菌株组β-内酰胺酶活性(12±6)U/mg比较,泛耐药菌株组β-内酰胺酶活性(253±36)U/mg,多重耐药菌株组β-内酰胺酶活性均明显增多,差异显著,有统计学意义(P<0.05)。而泛耐药菌株组β-内酰胺酶活性表达最高;与敏感菌株组细胞外膜通透性比较,泛耐药菌株组细胞外膜通透性、多重耐药菌株组细胞外膜通透性具有明显的阻碍作用,差异显著,有统计学意义(P<0.05)。而泛耐药菌株组细胞外膜通透性阻碍作用最强。结论β-内酰胺酶活性和细胞外膜通透性是肺炎克雷伯菌对β-内酰胺类抗生素耐药性的主决定因素。     

体外诱导多黏菌素耐药肺炎克雷伯菌的适合度代价相关研究

目的 研究体外诱导多黏菌素耐药肺炎克雷伯菌的适合度代价，并分析耐药株的分子特性。方法 采用体外诱导方法将临床分离的3株多黏菌素敏感肺炎克雷伯菌诱导成多黏菌素耐药株；微量肉汤稀释法检测体外诱导耐药前后菌株对临床常用抗菌药物的最低抑菌浓度(minimal inhibititory concentration, MIC)；采用PCR方法检测多黏菌素耐药相关基因pmrA、pmrB、mgrB、phoP和phoQ，并对阳性片段进行测序比对分析；对诱导耐药菌株及其对应敏感菌株进行生长曲线分析、生物膜形成能力检测及体外竞争和抗血清试验，分析研究多黏菌素耐药肺炎克雷伯菌的适合度代价。结果 3株诱导耐药株中均检出pmrB(T157P)突变，在诱导株FK713R和FK729R携带的mgrB基因中分别检出ISkpn14和IS5like插入序列。诱导耐药株与敏感株相比，24h生长曲线未发现明显差异；生物膜形成能力检测结果发现诱导耐药株与敏感株生物膜形成能力无明显差异；3株菌获得多黏菌素耐药后均表现出一定程度的体外竞争缺陷，竞争指数(CI值)分别为0.01、0.54和5×10-4；3株诱导耐药菌中有2株(FK713R和FK729R)与其敏感菌相比出现抗血清作用增强现象。结论 肺炎克雷伯菌获得多黏菌素耐药后会出现一定的适合度改变，不同的耐药机制可能会引起不同的适合度表现。     

Review of use of a new rapid real-time PCR,the Cepheid GeneXpert® (Xpert) CT/NG assay,for Chlamydia trachomatis and Neisseria gonorrhoeae: results for patients while in a clinical setting

Rapid diagnostics for Chlamydia trachomatis and Neisseria gonorrhoeae are desirable so that patients can be treated while they are still in the clinic or doctor’s office. The Cepheid GeneXpert^® (Xpert) CT/NG assay was US FDA-cleared in December 2012. The assay is a rapid real-time PCR nucleic acid amplified test. The cartridge-based assay detects DNA of Chlamydia trachomatis and Neisseria gonorrhoeae. It is FDA-cleared for use in female endocervical swabs, patient-collected vaginal swabs and for female and male urine specimens from symptomatic and asymptomatic patients. It has demonstrated near-perfect sensitivity and specificity in urogenital specimens. The Xpert is a modular platform for testing samples directly from patients, which requires no hands-on manipulation from specimen loading until results are available. Results are provided in approximately 90 minutes. It has been graded by the FDA as moderately complex for Clinical Laboratory Improvement Amendments. Several publications have reported its promising use in clinical settings.