摘要
目的检测SD大鼠腹腔感染前后血、粪纳米细菌(nanobacteria,NB),探讨NB与严重腹腔感染的相关性。方法40只SD大鼠,通过盲肠结扎加穿孔法制作SD大鼠严重腹腔感染模型,分别于感染前后取血、粪标本进行电镜下观察;NB培养;NB16SrRNA基因聚合酶链反应(polymerasechainreaction,PCR)扩增,电泳分析。结果电镜下观察NB,感染前、后血阳性率分别为17.5%和35.0%,P=0.118;感染前、后粪阳性率分别为40.0%和32.5%,P=0.250。经28d培养,结果均未见有NB特异性白色沉淀物形成。PCR法检测NB16SrRNA基因,感染前、后血阳性率分别为52.5%和90.0%,P=0.000;感染前、后粪阳性率均为95.0%。结论PCR法检测NB16SrRNA基因,SD大鼠粪普遍阳性,腹腔感染前、后血阳性率差异有显著意义,其作用尚待进一步研究。
Objective:To detect nanobacteria(NB)from blood and stool of rats suffering severe infection of ab-dominal cavity.Methods:40SD rats were involved in the study.The severe infection models of abdominal cavity were made by cecal ligation plus puncture(CLP).The test samples of blood and stool were obtained before or after infected respectively and filtered through0.22μm filters to eradicate the general germs and cells.The samples were stained with phosphoric tungstic acid and observed with transmission electron microscopy(TEM).RPMI1640culture with10.0%fetal bovine serum(FBS)was added into the filtered samples for culturing NB.Polymerase chain reaction(PCR)was made with the primers that designed from the sequences of NB16S rRNA gene recorded in the genebank(GenBank accession No.X98419) .PCR products were analyzed by agarose gel electrophoresis.Results:NB was detected with TEM from some samples.The positive rates before and after CLP respectively were17.5%and35.0%in the blood samples(P=0.118),40.0%and32.5%in the stool samples(P=0.250).We failed to find any white pre-cipitate in any culture bottle in28d.PCR assay was very sensitive for identify NB16S rRNA gene.The positive rates before and after CLP respectively were52.5%and90.0%in the blood samples(P=0.000),95.0%and95.0%in the stool samples(P=1.000).Conclusions:There exists NB in the stool of rats commonly by PCR assay and the difference is significant between the positive rates of the blood sample before and after CLP.It deserves to further insight on the relationship between NB and infection of abdominal cavity.
出处
《中国现代医学杂志》
CAS
CSCD
2004年第22期12-16,20,共6页
China Journal of Modern Medicine
