支原体感染状况调查及其分型以及致病机制的探讨
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摘要
目的:
应用解脲脲原体液体与固体双重培养法比较男性不育症患者与正常体检者解脲脲原体感染率的差异,并通过PCR的方法对解脲脲原体分群,探讨两组人群不同亚群解脲脲原体感染率的差异,同时对解脲脲原体培养阳性的不育症患者标本进行精液常规的检测,并利用激光共聚焦显微镜,观察免疫荧光定位的各群解脲脲原体感染状态及精子形态结构,从而探讨不同生物亚群解脲脲原体感染对精液常规及精子形态影响的差异,以便为临床诊断和治疗提供参考;通过PCR方法对天津地区非典型肺炎患儿肺炎支原体的感染率进行调查,同时收集该类患者七种病毒感染的情况,了解不同病原体感染率的差异,为丰富国内儿童肺炎支原体感染的流行病学研究状况提供资料,同时为指导临床治疗提供依据。
方法:
(1)选择2007年在天津市某三甲医院就诊的不育症患者75例,以及体检的正常男性270例,进行解脲脲原体的液体培养和固体培养,比较两组人群解脲脲原体阳性率的差异。
(2)提取阳性标本液体培养物的DNA,进行PCR分群,比较两组人群不同亚群解脲脲原体感染的差异。
(3)不育症患者的精液标本同时做精液常规以及用激光共聚焦显微镜观察免疫荧光定位的各群解脲脲原体感染状态、精子形态结构的情况。
(4)选择在天津市某儿童医院住院的患有非典型肺炎的患儿100例,采集其下呼吸道灌洗液标本,调查此类人群肺炎支原体的感染情况,同时收集该类患者七种病毒感染的情况,了解不同病原体感染率的差异。
结果:
(1)75例不育症患者精液标本经过液体培养后,47例阳性,阳性率为62.67%,液体培养阳性的标本转种固体培基后,42例阳性,最终阳性率为56%;270例正常体检男性精液标本经过液体培养后,78例阳性,阳性率为28.89%,液体阳性标本经过固体培养后,70例阳性,最终阳性率为25.93%。两组人群解脲脲原体感染阳性率的差异有统计学意义(x~2=24.213,P<0.05)。
(2)用PCR方法对42例解脲脲原体固体培养阳性的不育症患者及70例正常体检男性的Uu液体培养物进行分群,不育症患者Uu1群19例,占45.24%,2群18例,占42.86%,混合群5例,占11.9%;正常体检男性Uu1群51例,占72.86%,Uu2群17例,占24.29%,混合群2例,占2.86%,两组人群之间的差异有统计学意义(x~2=9.539,P<0.05)。
(3)解脲脲原体阳性的不育症患者标本精液常规结果显示,感染Uu1群与2群的患者标本之间,1群与混合群标本之间的存活率、活动率和畸形率的差异具有统计学意义(P<0.05),2群与混合群标本之间的差异无统计学意义(P>0.05)。
(4)激光共聚焦显微镜结果显示,解脲脲原体2群及混合群多定位于精子头部或尾部,而1群则多散在于精浆中,感染2群或混合群的不育症患者精子损伤情况明显比感染1群的患者严重。
(5)天津市某儿童医院非典型肺炎患儿100例,肺炎支原体的阳性率可达61%,而七种病毒包括人类呼吸道合胞病毒(RSV)、流行性感冒病毒A型(IFV A)、流行性感冒病毒B型(IFV B)、副流感病毒1型(PIV1)、副流感病毒2型(PIV2)、副流感病毒3型(PIV3)、腺病毒(ADV)的阳性率分别为31%、2%、0、0、0、4%、0,统计结果显示,肺炎支原体的检出率与七种病毒的检出率差异有统计学意义(P<0.05)。
结论:
(1)解脲脲原体液体培养存在一定的假阳性,临床上不能单纯以液体培养基颜色的变化来判定解脲脲原体阳性,应该转种固体培基以排除假阳性。
(2)本研究所做不育症患者与正常体检男性解脲脲原体的感染情况不同,不育症患者解脲脲原体阳性率明显高于正常男性,说明解脲脲原体感染与男性不育有一定的相关性。
(3)本研究中不育症患者与正常体检男性解脲脲原体亚群的感染情况不同,不育症患者Uu2群的感染率要比正常男性高,而正常体检者Uu1群的检出率高于不育症患者,说明Uu2群的致病性要高于Uu1群,Uu1群致病性较弱,或者为正常携带。
(4)Uu2群或混合群对精子的功能性或器质性的损伤高于Uu1群,而Uu2群和混合群之间的差异则无统计学意义,说明影响精子质量的主要是Uu2群。
(5)本研究所做天津地区非典型肺炎患儿肺炎支原体的感染率高于其他地区的检出率。
(6)天津地区非典型肺炎患儿肺炎支原体的检出率高于七种病毒的检出率,临床上应该高度重视肺炎支原体的检查,更好的指导临床用药。
Objective:
To compare the differences of Ureaplasma urealyticum infection rate between male infertility and normal male by liquid and solid culture.The Polymerase chain reaction was used to separate the Ureaplasma Urealyticum into two subtypes.The sperm routine examinations were applied in patients whose solid culture was positive and the Confocal laser scanning microscope was used to examine the condition of different subtypes of Ureaplasma Urealyticum infection and the morphology of the sperm.The Mycoplasma pneumonia infection rate of children who had atypical pneumonia in Tianjin was investigated by conventional PCR and confirmatory PCR. At the same time,the statuses of seven common virus infections in children were tested.And then,the infection rates of different pathogens were compared.Our study could offer data to clinical therapy and the epidemiology of Mycoplasma pneumoniae in China.
Methods:
(1) 75 cases of male infertility and 270 cases of normal male who went to a hospital of Tianjin in 2007 were enrolled in the study.And their sperm specimens were inoculated both in the liquid and solid medium.Ureaplasma Urealyticum infection rates were compared.
(2) DNA from those Ureaplasma Urealyticum liquid culture positive specimen was extracted.PCR was used to separate the two different subtype of Uu and the infection rate of them was compared.
(3) The sperm routine examination was taken and the Confocal laser scanning microscope was used to examine the condition of different subtypes of Ureaplasma Urealyticum infection.
(4) 100 cases of children who got atypical pneumonia in a children's hosipital of Tianjin were chosen to take part in the investigation of the Mycoplasma pneumoniae. BALF collecting from infected children were studied.And the infection rates of seven common viruses in children were collected.The infection rate of different pathogen were compared.
Results:
(1) 47 of the 75 cases of male infertility patients after the liquid culture of the sperm specimens were positive,and the positive rate was 62.67%.The positive cases of the solid culture were 42,and the final positive rate was 56%;The positive cases of the 270 cases of normal male after the liquid culture of the sperm specimens were 78, and the positive rate was 28.89%.And the positive cases of the solid culture were 70, and the final positive rate was 25.93%.The difference has statistical significance(x~2= 24.213,P＜0.05).
(2) Three subtypes of Ureaplasma Urealyticum was detected by PCR,in which biovar 1 was positive in 19 samples(45.24%),biovar 2 was positive in 18 samples(42.86%),and both biovar 1 and 2 positve was 5(11.9%) in 42 cases of male infertility;biovar 1 was positive in 51 samples(72.86%),biovar 2 was positive in 17 samples(24.29%),and both biovar 1 and 2 positve positive was 2(2.86%) in normal male.There was significant difference between the two groups(x~2=9.539,P＜0.05).
(3) It was shown by sperm routine examination that there's statistical significance between biovar 1 and biovar 2(P＜0.05),biovar 1 and mixed biovars(P＜0.05),but there wasn't statistical significance between bioval 2 and mixed biovar(P＞0.05) of the male infertility.
(4) It was also shown by the Confocal laser scanning microscope that the majority of biovar 2 and mixed biovars of Ureaplasma Urealyticum was in sperm head and tail, but many biovar 1 were free in the seminal plasma.The damage of biovar2 and mixed biovar was more severe than biovar 1.
(5) The Mycoplasma pneumoniae positive cases of children who got atypical pneumonia in 100 cases were 61(61%),but the positive rate of the seven kinds of virus,which containing Human respiratory syncytial virus(RSV)、Influenza A virus(IFVA)、Influenza B virus(IFVB)、Parain-fluenza virus 1(PIV1)、Parain-fluenza virus 2(PIV2)、Parain-fluenza virus 3(PIV3) and Adenovirus(ADV) were 31%、2%、0、0、0、4%and 0,respectively.The detection rate between the Mycoplasma pneumoniae and the seven virus has statistical difference(P＜0.05).
Conclusions:
(1) False positive results were found in the liquid culture of Ureaplasma urealyticum,and the color change doesn't always means Uu positive.The samples should be inoculated into the solid culture media to make a final decision.
(2) The infection situation of male infertility and normal male is different.The positive rate of male infertility is higher than normal male.It is concluded that there is some correlation between Uu infection and male infertility.
(3) The subtypes of Ureaplasma Urealyticum were different between patients with male infertility and normal male.The detection rate of biovar 2 and mixed biovar of Ureaplasma Urealyticum in patients with male infertility was higher than normal male.It is proved that the pathogenicity of biovar 2 and mixed biovars infection were severe than biovar 1.
(4) According to statistic results,biovar 2 caused more sever function and organic damage than biovar 1.So,the virulent of biovar 2 was strong.
(5) The infection rate of Mycoplasma pneumoniae in children with aptical pneumonia in Tianjin was higher than other areas in China.
(6) The detection rate of Mycoplasma pneumoniae is higher than seven kinds of virus.And we should pay more attention on Mycoplasma pneumonia detection for provident evidence to clinical practice.
应用解脲脲原体液体与固体双重培养法比较男性不育症患者与正常体检者解脲脲原体感染率的差异,并通过PCR的方法对解脲脲原体分群,探讨两组人群不同亚群解脲脲原体感染率的差异,同时对解脲脲原体培养阳性的不育症患者标本进行精液常规的检测,并利用激光共聚焦显微镜,观察免疫荧光定位的各群解脲脲原体感染状态及精子形态结构,从而探讨不同生物亚群解脲脲原体感染对精液常规及精子形态影响的差异,以便为临床诊断和治疗提供参考;通过PCR方法对天津地区非典型肺炎患儿肺炎支原体的感染率进行调查,同时收集该类患者七种病毒感染的情况,了解不同病原体感染率的差异,为丰富国内儿童肺炎支原体感染的流行病学研究状况提供资料,同时为指导临床治疗提供依据。
方法:
(1)选择2007年在天津市某三甲医院就诊的不育症患者75例,以及体检的正常男性270例,进行解脲脲原体的液体培养和固体培养,比较两组人群解脲脲原体阳性率的差异。
(2)提取阳性标本液体培养物的DNA,进行PCR分群,比较两组人群不同亚群解脲脲原体感染的差异。
(3)不育症患者的精液标本同时做精液常规以及用激光共聚焦显微镜观察免疫荧光定位的各群解脲脲原体感染状态、精子形态结构的情况。
(4)选择在天津市某儿童医院住院的患有非典型肺炎的患儿100例,采集其下呼吸道灌洗液标本,调查此类人群肺炎支原体的感染情况,同时收集该类患者七种病毒感染的情况,了解不同病原体感染率的差异。
结果:
(1)75例不育症患者精液标本经过液体培养后,47例阳性,阳性率为62.67%,液体培养阳性的标本转种固体培基后,42例阳性,最终阳性率为56%;270例正常体检男性精液标本经过液体培养后,78例阳性,阳性率为28.89%,液体阳性标本经过固体培养后,70例阳性,最终阳性率为25.93%。两组人群解脲脲原体感染阳性率的差异有统计学意义(x~2=24.213,P<0.05)。
(2)用PCR方法对42例解脲脲原体固体培养阳性的不育症患者及70例正常体检男性的Uu液体培养物进行分群,不育症患者Uu1群19例,占45.24%,2群18例,占42.86%,混合群5例,占11.9%;正常体检男性Uu1群51例,占72.86%,Uu2群17例,占24.29%,混合群2例,占2.86%,两组人群之间的差异有统计学意义(x~2=9.539,P<0.05)。
(3)解脲脲原体阳性的不育症患者标本精液常规结果显示,感染Uu1群与2群的患者标本之间,1群与混合群标本之间的存活率、活动率和畸形率的差异具有统计学意义(P<0.05),2群与混合群标本之间的差异无统计学意义(P>0.05)。
(4)激光共聚焦显微镜结果显示,解脲脲原体2群及混合群多定位于精子头部或尾部,而1群则多散在于精浆中,感染2群或混合群的不育症患者精子损伤情况明显比感染1群的患者严重。
(5)天津市某儿童医院非典型肺炎患儿100例,肺炎支原体的阳性率可达61%,而七种病毒包括人类呼吸道合胞病毒(RSV)、流行性感冒病毒A型(IFV A)、流行性感冒病毒B型(IFV B)、副流感病毒1型(PIV1)、副流感病毒2型(PIV2)、副流感病毒3型(PIV3)、腺病毒(ADV)的阳性率分别为31%、2%、0、0、0、4%、0,统计结果显示,肺炎支原体的检出率与七种病毒的检出率差异有统计学意义(P<0.05)。
结论:
(1)解脲脲原体液体培养存在一定的假阳性,临床上不能单纯以液体培养基颜色的变化来判定解脲脲原体阳性,应该转种固体培基以排除假阳性。
(2)本研究所做不育症患者与正常体检男性解脲脲原体的感染情况不同,不育症患者解脲脲原体阳性率明显高于正常男性,说明解脲脲原体感染与男性不育有一定的相关性。
(3)本研究中不育症患者与正常体检男性解脲脲原体亚群的感染情况不同,不育症患者Uu2群的感染率要比正常男性高,而正常体检者Uu1群的检出率高于不育症患者,说明Uu2群的致病性要高于Uu1群,Uu1群致病性较弱,或者为正常携带。
(4)Uu2群或混合群对精子的功能性或器质性的损伤高于Uu1群,而Uu2群和混合群之间的差异则无统计学意义,说明影响精子质量的主要是Uu2群。
(5)本研究所做天津地区非典型肺炎患儿肺炎支原体的感染率高于其他地区的检出率。
(6)天津地区非典型肺炎患儿肺炎支原体的检出率高于七种病毒的检出率,临床上应该高度重视肺炎支原体的检查,更好的指导临床用药。
Objective:
To compare the differences of Ureaplasma urealyticum infection rate between male infertility and normal male by liquid and solid culture.The Polymerase chain reaction was used to separate the Ureaplasma Urealyticum into two subtypes.The sperm routine examinations were applied in patients whose solid culture was positive and the Confocal laser scanning microscope was used to examine the condition of different subtypes of Ureaplasma Urealyticum infection and the morphology of the sperm.The Mycoplasma pneumonia infection rate of children who had atypical pneumonia in Tianjin was investigated by conventional PCR and confirmatory PCR. At the same time,the statuses of seven common virus infections in children were tested.And then,the infection rates of different pathogens were compared.Our study could offer data to clinical therapy and the epidemiology of Mycoplasma pneumoniae in China.
Methods:
(1) 75 cases of male infertility and 270 cases of normal male who went to a hospital of Tianjin in 2007 were enrolled in the study.And their sperm specimens were inoculated both in the liquid and solid medium.Ureaplasma Urealyticum infection rates were compared.
(2) DNA from those Ureaplasma Urealyticum liquid culture positive specimen was extracted.PCR was used to separate the two different subtype of Uu and the infection rate of them was compared.
(3) The sperm routine examination was taken and the Confocal laser scanning microscope was used to examine the condition of different subtypes of Ureaplasma Urealyticum infection.
(4) 100 cases of children who got atypical pneumonia in a children's hosipital of Tianjin were chosen to take part in the investigation of the Mycoplasma pneumoniae. BALF collecting from infected children were studied.And the infection rates of seven common viruses in children were collected.The infection rate of different pathogen were compared.
Results:
(1) 47 of the 75 cases of male infertility patients after the liquid culture of the sperm specimens were positive,and the positive rate was 62.67%.The positive cases of the solid culture were 42,and the final positive rate was 56%;The positive cases of the 270 cases of normal male after the liquid culture of the sperm specimens were 78, and the positive rate was 28.89%.And the positive cases of the solid culture were 70, and the final positive rate was 25.93%.The difference has statistical significance(x~2= 24.213,P＜0.05).
(2) Three subtypes of Ureaplasma Urealyticum was detected by PCR,in which biovar 1 was positive in 19 samples(45.24%),biovar 2 was positive in 18 samples(42.86%),and both biovar 1 and 2 positve was 5(11.9%) in 42 cases of male infertility;biovar 1 was positive in 51 samples(72.86%),biovar 2 was positive in 17 samples(24.29%),and both biovar 1 and 2 positve positive was 2(2.86%) in normal male.There was significant difference between the two groups(x~2=9.539,P＜0.05).
(3) It was shown by sperm routine examination that there's statistical significance between biovar 1 and biovar 2(P＜0.05),biovar 1 and mixed biovars(P＜0.05),but there wasn't statistical significance between bioval 2 and mixed biovar(P＞0.05) of the male infertility.
(4) It was also shown by the Confocal laser scanning microscope that the majority of biovar 2 and mixed biovars of Ureaplasma Urealyticum was in sperm head and tail, but many biovar 1 were free in the seminal plasma.The damage of biovar2 and mixed biovar was more severe than biovar 1.
(5) The Mycoplasma pneumoniae positive cases of children who got atypical pneumonia in 100 cases were 61(61%),but the positive rate of the seven kinds of virus,which containing Human respiratory syncytial virus(RSV)、Influenza A virus(IFVA)、Influenza B virus(IFVB)、Parain-fluenza virus 1(PIV1)、Parain-fluenza virus 2(PIV2)、Parain-fluenza virus 3(PIV3) and Adenovirus(ADV) were 31%、2%、0、0、0、4%and 0,respectively.The detection rate between the Mycoplasma pneumoniae and the seven virus has statistical difference(P＜0.05).
Conclusions:
(1) False positive results were found in the liquid culture of Ureaplasma urealyticum,and the color change doesn't always means Uu positive.The samples should be inoculated into the solid culture media to make a final decision.
(2) The infection situation of male infertility and normal male is different.The positive rate of male infertility is higher than normal male.It is concluded that there is some correlation between Uu infection and male infertility.
(3) The subtypes of Ureaplasma Urealyticum were different between patients with male infertility and normal male.The detection rate of biovar 2 and mixed biovar of Ureaplasma Urealyticum in patients with male infertility was higher than normal male.It is proved that the pathogenicity of biovar 2 and mixed biovars infection were severe than biovar 1.
(4) According to statistic results,biovar 2 caused more sever function and organic damage than biovar 1.So,the virulent of biovar 2 was strong.
(5) The infection rate of Mycoplasma pneumoniae in children with aptical pneumonia in Tianjin was higher than other areas in China.
(6) The detection rate of Mycoplasma pneumoniae is higher than seven kinds of virus.And we should pay more attention on Mycoplasma pneumonia detection for provident evidence to clinical practice.
引文
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