Diagnosis Optimization Of Human Eperythrozoonsis And In Vitro Culture Of Human Eperythrozoon

Eperythrozoon(EPE)is a kind of conditional pathogenic microorganism.The proportion of human infection with eperythrozoon is increasing year by year.The harm of eperythrozoon to human body has gradually attracted the attention of the medical community.At present,there are several methods for eperythrozoon detection such as direct microscopy,staining microscopy and PCR in the laboratory.In this topic we did the survey on the eperythrozoon infection rates of patients with blood diseases,detected eperythrozoon in man by a variety of combined methods,and established the in-vitro cultivation of human eperythrozoon preliminarily.1.We used acridine orange fluorescence staining to investigate the infection of eperythrozoon in the patients with blood disease so as to provide evidence to control the infection of the patients with blood disease.A total of 50 healthy subjects and 223 patients with blood diseases were enrolled.Eperythrozoon was determined by acridine orange-stained.The results showed the infection rates in healthy subject group and blood disease group by acridine orange-stained blood smears were 12.00% and 28.25%,respectively.The infection rate in blood disease group was higher than that in healthy subject group(?2=5.711,P<0.05).No difference of infection rate was found between males and females(P>0.05).The infection rates showed no statistical significance for ages(P>0.05).The infection rates between outpatients from hematology department and inpatients had no statistical significance(P>0.05).The infection rate of patients with lymphoma was the highest in blood disease group.There was statistical significance for the infection rate between patients with lymphoma and patients with other blood diseases(P<0.05).The result showed that patients with blood diseases especially with lymphoma are more susceptible than healthy subjects to eperythrozoon.Thus,attention should be paid during treatment.2.223 patients with blood diseases were enrolled.These samples were respectively detected through Wright-Giemsa blood smearsand acridine orange-stained blood smears.It was found that the positive rates of 223 patients with blood diseases were 46.19% and 28.25% by Wright-Giemsa blood smears and acridine orange-stained blood smears,respectively.40 patients with both positive results were expected to do PCR tests.One of the positive results was sequenced and analyzed.The sequence was 97% similar to that of Mycoplasma suisaccording to GenBank.The results showed Wright-Giemsa blood smears and(or)acridine orange-stained blood smears could be used for screening eperythrozoon infection.When both methods were positive,PCR detection could improve the specificity of eperythrozoon detection.3.RPMI-1640 medium and fetal bovine serum were mixed in a certain proportion,and glucose was added as the basic medium.Normal human red blood cell mud was used as the carrier,and inoculated red blood cells infected with human eperythrozoon for in-vitro cultivation.Genomic DNA was extracted from the culture material for PCR detection and analysis.The proliferation peak of human eperythrozoon was observed at 24 h in RPMI-1640 culture medium with normal erythrocytes,and the infection rate was about 80%.The results of PCR detection were consistent with the sequence of human eperythrozoon genome extracted from the patient's original blood sample.In this study,we further analyzed and compared the laboratory detection system of eperythrozoon in human,and initially established the culture technology of human eperythrozoonsis in vitro,which provided theoretical basis and experimental basis for further study of human eperythrozoonsis in the future.