COMPARISON OF 3 METHODS FOR EXTRACTION OF VIRAL NUCLEIC-ACIDS FROM BLOOD CULTURES
COMPARISON OF 3 METHODS FOR EXTRACTION OF VIRAL NUCLEIC-ACIDS FROM BLOOD CULTURES
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DOI：
10.1128/jcm.33.1.41-44.1995
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发表时间：
1995-01-01
影响因子：
9.4
通讯作者：
SMITH, TF
中科院分区：
文献类型：
--
作者：
ESPY, MJ;PATEL, R;SMITH, TF
为了灵敏地检测病毒DNA，需要可靠的血液标本核酸提取技术。处理用于疱疹病毒分子检测的血液标本的标准化程序（巨细胞病毒[CMV]、单纯疱疹病毒、水痘-带状疱疹病毒和爱泼斯坦-巴尔病毒[EBV]）尚未建立。使用三种方法从来自健康供体和无症状免疫功能低下患者的血液标本中提取DNA：（i）IsoQuick处理全血，（ii）通过裂解提取外周血白细胞（裂解缓冲液和蛋白酶K），和（iii）用苯酚-氯仿提取外周血白细胞（十二烷基硫酸钠溶液和蛋白酶K），来自25名健康献血者的所有血液标本均为CMV、单纯疱疹病毒和水痘-带状疱疹病毒核酸序列阴性，无论提取方法如何，而通过裂解技术提取的三个样品（12%）对EBV DNA呈阳性。在25例无症状免疫功能低下患者的血液标本中，用裂解提取法检测了9例CMV和EBV，用IsoQuick法检测了4例CMV和6例EBV，用酚-氯仿法检测了4例CMV和6例EBV。这些病毒的DNA经常存在于无症状免疫功能低下患者的血液标本中，偶尔也存在于健康供体的血液标本中。
Reliable nucleic acid extraction techniques for blood specimens are required for the sensitive detection of viral DNA. Standardized procedures for processing blood specimens for the molecular detection of herpesviruses (cytomegalovirus [CMV], herpes simplex virus, varicella-zoster virus, and Epstein-Barr virus [EBV]) have not been established, Three methods were used to extract DNA from blood specimens from healthy donors and asymptomatic immunocompromised patients: (i) IsoQuick treatment of whole blood, (ii) extraction of the peripheral blood leukocytes by lysis (lysis buffer and proteinase K), and (iii) extraction of peripheral blood leukocytes with phenol-chloroform (sodium docecyl sulfate solution and proteinase K), All blood specimens from 25 healthy blood donors were negative for CMV, herpes simplex virus and varicella-zoster virus nucleic acid sequences, regardless of the extraction method, while three samples (12%) extracted by the lysis technique were positive for EBV DNA. Of 25 blood samples from asymptomatic immunocompromised patients, CMV and EBV each were detected in nine specimens by lysis extraction, four each by IsoQuick and four (CMV) and six (EBV) by the phenol-chloroform method, Our results indicate that the lysis method is optimal for the detection of CMV and EBV DNA sequences by PCR from the leukocytic fraction of blood specimens. DNA of these viruses is frequently present in blood specimens from asymptomatic immunocompromised patients and occasionally from healthy donors.