A 41-kDa variable surface protein of Mycoplasma gallisepticum has a counterpart in Mycoplasma imitans and Mycoplasma iowae

Abstract Mycoplasma gallisepticun, M. imitans and M. iowae are three morphologically similar avian Mycoplasma species, and M. gallisepticum and M. imitans have been shown to be antigenically related. Using a monoclonal antibody that binds to the previously described size- and phase-variant integral membrane surface protein PvpA of M. gallisepticum , we have identified in all three avian Mycoplasma species a 41-kDa surface antigen, which in M. gallisepticum and M. imitans was identified as peripheral membrane protein undergoing variation in expression among clonal isolates. Southern blot analysis using the pvpA gene as a probe demonstrated sequence homology with M. imitans and M. iowae genomic DNA and suggested that a pvpA -related gene that may encode the 41-kDa product exists in these two Mycoplasma species. These studies establish (i) that M. iowae is antigenically related to M. gallisepticum and M. imitans , (ii) that the three species share non-ribosomal gene sequences, and (iii) that peripheral membrane proteins contribute to Mycoplasma surface variation.     

Biochemical diversity of Mycoplasma fermentans strains

In this study, the metabolism of a diverse range of Mycoplasma fermentans strains was investigated. It was shown that the ability to utilise glucose, fructose and N-acetylglucosamine differentiated strains, and that the patterns and kinetics of substrate utilisation were correlated with the site of isolation, i.e. joint fluid, respiratory tract, urinary tract or cell culture. Interestingly, isolates from the urogenital tract of AIDS patients used fructose in preference to glucose. There was also some correlation of fructose and N-acetylglucosamine utilisation of isolates with M. fermentans sub-groups, identified in an independent study, and based on the distribution of insertion sequence-like elements in the M. fermentans genome.     

猪肺炎支原体及其他支原体黏附因子的研究进展

猪肺炎支原体能够引起猪支原体肺炎,其黏附因子在致病过程中起重要作用。本文综合国内外猪肺炎支原体黏附因子的研究进展,并与其他支原体的黏附因子进行了比较讨论,从而为猪肺炎支原体致病机理的进一步研究及该病的防制提供新思路。     

细胞培养物中污染支原体的去除

经小鼠体内、体外加用不同的抗菌素处理及克隆的方法处理细胞污染的支原体,三株细胞经两次、一株细胞经三次处理后,经培养法、ＤＮＡ染色法及ＰＣＲ法检查支原体污染均为阴性,并证明对其抗体分泌没有任何影响,不失为一个理想的去除支原体的方法,从而使有重要应用价值的支原体污染细胞的应用成为可能。     

女性生殖道炎症支原体多重耐药性分析

了解本地区在采用常规抗生素治疗支原体后多重耐药株的产生情况,为临床针对性用药提供科学依据。采用培养法对女性生殖道炎患者分泌物进行支原体培养、体外药敏试验及分析。结果可见,感染数量≥10~4 cfu/mL的支原体阳性率为52.9%(82/155);13种抗生素中,耐药种数构成比为6R>2R、4R、5R>3R>1R>8R>7R、10R>0R>9R、12R、13R;耐药率为林可霉素>壮观霉素>环丙沙星>诺氟沙星>司帕沙星>罗红霉素>氧氟沙星>克拉霉素>左氧氟沙星>阿齐霉素>强力霉素,美满霉素>交沙霉素。体外药敏试验对临床治疗支原体感染及减少多重耐药株的产生具有重要指导意义。     

11788例儿童血清肺炎支原体抗体检测结果分析

目的了解宁波儿童肺炎支原体（MP）感染情况,为儿童MP感染的临床诊断与防治提供依据。方法采用被动凝集法测定11788例14岁以下有呼吸道感染症状儿童的血清MP抗体,并根据患儿性别、年龄和不同季节对MP的阳性率进行统计学分析。结果11788例有呼吸道感染症状患儿中,MP阳性3081例,阳性率为26．14％,不同性别、年龄和季节MP抗体的阳性率差异有统计学意义（P〈0．05）。女性儿童阳性率高于男性儿童,并随着年龄的增加阳性率有上升趋势。结论MP在呼吸道感染中发病率较高,春冬季节、女性儿童和学龄期儿童易感。     

Tetrazolium reduction methods for assessment of substrate oxidation and strain differentiation among mycoplasmas, with particular reference to Mycoplasma bovigenitalium and some members of the Mycoplasma mycoides cluster

Aims:  To apply a rapid nitroblue tetrazolium (NBT) reduction assay of substrate metabolism by mycoplasmas that would help to differentiate Mycoplasmas .
Methods and Results:  Growth, substrate preferences and tetrazolium reduction were assessed for 18 strains of Mycoplasma bovigenitalium and Mycoplasma ovine serogroup 11. NBT reduction was detectable in 1 h with 10⁸ CFU ml⁻¹. Use of α-ketobutyrate, lactate and pyruvate to support growth and NBT reduction were correlated: pyruvate was preferred and lactate was used by only four of the 18 strains. Selected members of the Mycoplasma mycoides cluster were also assessed and monotetrazoles tested as alternatives to NBT. The NBT method was applied to a further 19 species.
Conclusions:  This simple and reproducible method requires only small amounts of cells, enabling routine assessment of substrate use within 1 h, and the rapid assignment of numerous mycoplasmas to one of six physiological groups. The four physiological groups of M. bovigenitalium and Mycoplasma serogroup 11 strains were indistinguishable from each other, which supports the view that these belong to the same species.
Significance and Impact of the Study:  Strain-specific substrate-utilization patterns by mycoplasmas can be obtained rapidly and reliably. The method has potential as a large-scale semi-automated procedure to monitor numerous strains and substrates simultaneously.     

支原体菌株低温保存活力稳定性研究

目的研究低温冻存时间及温度对支原体菌种保存稳定性和菌种活力的影响。方法用固体平板计数法计算不同冻存时间、温度下的口腔支原体、肺炎支原体菌落数差异,分析这2个因素对口腔支原体、肺炎支原体菌种活力的影响。结果口腔支原体在生长48 h后达到生长高峰,肺炎支原体生长较为缓慢,在103 h到达生长高峰。口腔支原体在-70℃冻存1个月时,菌落数由10¹² cfu/mL下降至10¹¹ cfu/mL,冻存1个月的菌落数和冻存3个月、12个月的菌落数比较,差异均无统计学意义(P>0.05)。肺炎支原体在-70℃的冻存条件下,菌落数一直处于稳定水平;比较-35℃与-70℃的冻存条件下肺炎支原体的菌落数,差异无统计学意义(P>0.05),口腔支原体在-70℃冻存条件下的菌种活力明显高于-35℃(P<0.05)。结论口腔支原体生长迅速,但对于储存温度的敏感性较高,肺炎支原体相较口腔支原体生长缓慢,停滞期较长,菌株对冻存温度和时间的敏感度较低,稳定性高。菌种冻存温度应选择较低温度,冻存周期在1年内较为稳定。     

Isolation and characterization of transposon Tn4001-generated, cytadherence-deficient transformants of Mycoplasma pneumoniae and Mycoplasma genitalium

Abstract Cytadherence and subsequent parasitism of host cells by the human pathogens, Mycoplasma pneumoniae and Mycoplasma genitalium , are mediated by adhesins and adherence-related accessory proteins. In this report we demonstrate the use of transposon Tn 4001 to generate Tn-induced transformants displaying cytadherence-deficient characteristics. Mycoplasma pneumoniae Tn-generated transformant, designated 8R, lacked the high-molecular weight adherence-accessory proteins HMW1/4 and was deficient in hemadsorption and cytadherence capabilities. In transformant 8R, Tn 4001 was not localized in or near the hmw 1 gene or in the upstream adhesin (p30/hmw3) locus, suggesting an alternate site associated with the regulation of hmw 1 gene expression. Sequence analysis identified the transposon insertion site at the crl locus previously reported, although the protein characteristics of transformant 8R differed from the earlier described transformants. The M. genitalium Tn-transformant, designated G26, was also defective in hemadsorption and cytadherence. However, transformant G26 synthesized adhesins P140 and P32 suggesting that Tn 4001 transposed into a new gene or site previously unlinked to cytadherence, namely ORF MG032. This study demonstrates the utility of Tn 4001 mutagenesis for both M. pneumoniae and M. genitalium which, in the latter case, has special relevance in light of the recent complete characterization of its continuous total genomic sequence.     

肺炎支原体实验室检测方法研究进展

肺炎支原体(Mycoplosma pneumonia,MP)为人类非典型肺炎的病原体,是引起呼吸道感染的重要病原体。但是支原体肺炎与其他病原体感染的肺炎,在临床症状、影像学上并无特异性差别,且其对一般治疗肺炎、上呼吸道感染的药物有耐药性,因此肺炎支原体及时、准确的实验室检测对于支原体肺炎的诊断治疗显得尤为重要。目前MP的实验室检测方法不断推陈出新,但各种方法均有其优势与不足,临床可选择两种不同的方法同时检测。比如:血清学抗体的检测结合MP快速培养药敏的方法;血清学抗体的检测结合PCR的方法,不同方法相互补充为临床的早期诊断、治疗提供依据。而MP药敏试验的检测和耐药机制的研究对于临床用药方案的选择,减少耐药株的产生和流行具有重要意义。     

Alternatives to arginine as energy sources for the non-fermentative Mycoplasma gallinarum

Abstract In contrast to previously studied non-fermentative arginine-hydrolysing (F-/A+) Mycoplasma species, M. gallinarum cells suspended in salts solution oxidised ethanol and L-lactic, pyruvic and 2-oxobutyric acids. The organic acids were additionally shown effectively to replace arginine as energy sources in growth media. However, their presence did not inhibit arginine hydrolysis, nor did arginine inhibit organic acid catabolism. The ability to oxidase organic acids is a potentially useful diagnostic character enabling sub-division of the F-/A+ Mycoplasma species. M. gallinarum also differed from previously studied F-/A+ mycoplasmas in possessing relatively high NADH oxidase activity and producing H₂O₂ as only a minor product of NADH oxidation.     

Distribution of ecto 5'-nucleotidase on Mycoplasma species associated with arthritis

The enzyme ecto 5'-nucleotidase (5'N) was found to be active on 8/14 strains of Mycoplasma fermentans, K(m) (+/-S.D.) 3.8+/-2.8 microM 5'-AMP, and on the type strain of Mycoplasma pulmonis, K(m) 0.63 microM 5'-AMP. The six M. fermentans strains lacking 5'N activity were related by restriction fragment length polymorphism typing. At pH 8.5, the type strains of Mycoplasma arthritidis, Mycoplasma buccale and Ureaplasma urealyticum showed a relatively non-specific phosphatase activity against 5'-AMP but no activity was shown by the type strains of Mycoplasma genitalium, Mycoplasma hominis, Mycoplasma orale, Mycoplasma penetrans, Mycoplasma pneumoniae and Mycoplasma salivarium at this pH. M. fermentans has been reported from rheumatoid joints, which show a raised 5'N activity on their synovial cells and in their fluid which may be associated directly or indirectly with the mycoplasma.     

Membrane lipids of Mycoplasma fermentans

Abstract Membranes of Mycoplasma fermentans , incognitus strain, were isolated by a combination of osmotic lysis and sonication. Analysis of membrane lipids revealed, in addition to free and esterified cholesterol, six major polar lipids dominated by a de novo synthesized compound (compound X), which accounts for 64% of the total lipid phosphorus. Compound X was labeled by palmitate, but not by oleate. Mass spectrometry and gas liquid chromatography analyses of compound X revealed two molecular species with molecular masses of 1048 and 1076 representing, a dipalmitoyl- and a stearoyl-palmitoyl-glycerodiphosphatidylcholine. Compound X has the ability to stimulate human monocytes to secret TNFα and to enhance the fusion of small unilamellar vesicles with MOLT-3 lymphocytes.     

常见支原体感染误诊与对策

临床支原体感染易发生误诊,不但加重患者精神和经济负担,而且延误患者病情,甚至留下后遗症。为此,我们综述了常见支原体感染误诊的特点和原因,深挖其中思维和思想根源,探讨减少或避免该类误诊现象的策略,旨在为临床医护人员和检验技术人员提供参考建议。     

小儿重症支原体肺炎高危因素分析

目的:筛选并分析重症支原体肺炎患儿的临床危险因素,为重症支原体肺炎的预防、诊断、治疗提供科学参考。方法:回顾性分析我院自2009年10月~2013年7月期间,确诊为支原体肺炎的患儿347例,根据患儿病情严重程度分为重症组和一般组,重症组有重症支原体肺炎患儿107例,一般组有患儿240例,从两组患儿的病例资料中抽取年龄、性别、居住地、(C reactive protein,CRP)、人肺炎支原体抗体(Mycoplasma pneumoniae antibody,MP-Ab)阳性率、病程、生活环境、反复呼吸道感染(Recurrent respiratory tract infection,RRI)、超敏C反应蛋白(High sensitivity C reactive protein,HSCRP)、红细胞沉积率(Erythrocyte sedimentation rate,ESR)、白细胞数(white blood cell,WBC)等相关指标,进行相关统计分析。对其分类指标进行合理赋值处理,实施单因素分析和多因素非条件的Logistic分析。结果:单因素分析显示:年龄、RRI、特殊体质、HSCRP、ESR以及先天性心脏病与重症支原体肺炎的发生相关(P=0.000,0.000,0.010,0.000,0.000,0.022);多因素非条件Logistic分析显示:年龄、RRI、HSCRP、ESR为重症支原体肺炎的危险因素(P=0.001,0.000,0.008,0.000)。结论:年龄、RRI、HSCRP、ESR是重症支原体肺炎的危险因素,通过对以上危险因素的密切关注对于重症支原体肺炎的预防、诊断、治疗有着重要的意义。     

儿科呼吸道疾病中肺炎支原体感染的结果分析

目的探讨儿科肺炎支原体（MP）感染特点,辅助临床医师早期诊断,合理用药。方法测定我院一年来2013例儿科呼吸道疾病患儿的肺炎支原体抗体（IgM）。结果2013例呼吸道感染患儿,检出肺炎支原体抗体（IgM）阳性者769例,占38.2%,769例阳性分别表现为肺炎369例（48%）,支气管炎238例（31%）,咽炎92例（12%）,哮喘70例（9%）。其中,肺炎组与各组相比较,具有统计学意义。MP IgM感染的检出率明显高于其他各组（P〈0.01）。结论MP感染是患儿不可忽视的病原体,检测患儿血清MP抗体能够及早诊断,指导治疗。     

PCR技术在鼠肺支原体检测中应用的初步研究

目的　探讨PCR技术在鼠肺支原体检测中的应用,希望能建立一种可行、快速、敏感的检测方法。方法　使用支原体通用引物及鼠肺支原体特异性引物对14 份大鼠喉气管拭子洗液和拭子支原体培养液进行PCR扩增,2 % 琼脂糖电泳鉴定。另设M53 和ATCC19612 二株标准鼠肺支原体菌株作阳性对照。结果　通用引物对大鼠喉气管拭子洗液检出率8/14 ,拭子支原体培养液检出率14/14,鼠肺支原体特异引物PCR扩增对大鼠喉气管拭子洗液检出率0/14 ,拭子支原体培养液3/14。通用引物扩增M53 和ATCC19612 二株标准株均呈现阳性,而鼠肺支原体特异引物扩增M53 和ATCC19612,只有M53 呈现阳性。结论　PCR通用引物检测比普通分离培养省时省力,而我们采用国外某学者认为对鼠肺支原体有特异性的引物,是否可用于鼠肺支原体的特异性PCR 检查仍需进一步探讨。     

非淋菌性泌尿生殖道炎支原体感染及耐药变迁

目的探讨解脲支原体（Uu）及人型支原体（Mh）在非淋菌性泌尿生殖道炎的感染状况,分析近3年来支原体的感染及耐药变迁。方法采用培养法（Mycoplasma IST）对NGGU的分泌物进行支原体的培养、体外药敏试验及分析。结果支原体阳性率为47．0％,其中Uu阳性率为34．0％,Uu＋Mh阳性率为11．4％,Mh阳性率为1．6％;交沙霉素、强力霉素等药物的敏感率无明显变化,而喹诺酮类的耐药率则呈逐渐上升趋势,环丙沙星及氧氟沙星的耐药率分别从2002年的38．9％及40．0％上升到了2004年的79．2％及66．4％。结论支原体的耐药率正逐步增加,其中喹诺酮类的高耐药率提示其在治疗支原体感染中的地位改变,泌尿生殖道支原体的药物敏感监测对临床治疗具有重要意义。