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恙虫病东方体在浓缩红细胞中的复苏与活力以及输血感染恙虫病的风险

Recovery and viability of Orientia tsutsugamushi from packed red cells and the danger of acquiring scrub typhus from blood transfusion.

作者信息

Casleton B G, Salata K, Dasch G A, Strickman D, Kelly D J

机构信息

Department of Clinical Investigation, Walter Reed Army Medical Center, Washington, DC, USA.

出版信息

Transfusion. 1998 Jul;38(7):680-9. doi: 10.1046/j.1537-2995.1998.38798346638.x.

Abstract

BACKGROUND

The purpose of this study was to determine whether infective Orientia tsutsugamushi, the etiologic agent of scrub typhus, could survive normal blood banking processing and storage procedures.

STUDY DESIGN AND METHODS

Mononuclear cells isolated from whole blood by density gradient centrifugation were inoculated with O. tsutsugamushi, Karp strain. Infection of the mononuclear cells was confirmed by Giemsa stain, direct fluorescent antibody assay, and polymerase chain reaction using primers specific for the groESL operon of O. tsutsugamushi. The quantity of rickettsial particles in each preparation was determined by direct counts from the Giemsa-stained preparations. Infected mononuclear cells were returned to their respective aliquots of packed red blood cells, which were then either stored at 4 degrees C or glycerolized and frozen at -70 degrees C.

RESULTS

Rickettsiae survived up to 10 days (but not 30 days) of refrigerated storage and 45 days of frozen storage, as determined by inoculation of mice with 0.5-mL aliquots of the blood components. Infection of the mice was determined by illness, death, direct fluorescent antibody assay of peritoneal smears, polymerase chain reaction of blood, and enzyme-linked immunosorbent assay detection of antibodies in plasma.

CONCLUSION

Because the quantity of rickettsiae injected into the mice was comparable to the quantity reported in the literature for human blood during natural infections, scrub typhus could present a risk in blood collected from donors in endemic areas. This may especially be true, because people can be rickettsemic before illness, after successful antibiotic treatment, and chronically after resolution of disease.

摘要

背景

本研究的目的是确定恙虫病东方体(丛林斑疹伤寒的病原体)能否在正常的血液储存处理程序中存活。

研究设计与方法

通过密度梯度离心从全血中分离出的单核细胞接种恙虫病东方体Karp株。用吉姆萨染色、直接荧光抗体检测以及使用针对恙虫病东方体groESL操纵子的特异性引物进行聚合酶链反应来确认单核细胞的感染情况。通过对吉姆萨染色制剂直接计数来确定每份制剂中立克次体颗粒的数量。将感染的单核细胞放回各自的红细胞成分等分试样中,然后将其在4℃储存或甘油化并在-70℃冷冻。

结果

通过用0.5 mL血液成分等分试样接种小鼠来确定,立克次体在冷藏储存10天(但不是30天)和冷冻储存45天的情况下仍能存活。通过疾病、死亡情况、腹膜涂片的直接荧光抗体检测、血液的聚合酶链反应以及血浆中抗体的酶联免疫吸附测定来确定小鼠是否感染。

结论

由于注入小鼠体内的立克次体数量与文献报道的自然感染期间人体血液中的数量相当,丛林斑疹伤寒可能会在流行地区从献血者采集的血液中构成风险。尤其可能如此,因为在患病前、成功抗生素治疗后以及疾病消退后慢性期人们都可能出现立克次体血症。

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