STORAGE AND PRESERVATION OF WHOLE-BLOOD SAMPLES FOR USE IN DETECTION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 BY THE POLYMERASE CHAIN-REACTION

被引:26
作者
KAYE, S
LOVEDAY, C
TEDDER, RS
机构
[1] Division of Virology, University College, Middlesex School of Medicine, London
关键词
HIV PCR; SAMPLE PREPARATION; DNA EXTRACTION;
D O I
10.1016/0166-0934(91)90137-O
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Methods used in the diagnosis of human immunodeficiency virus type-1 (HIV-1) infection by the polymerase chain reaction (PCR) usually require the separation of lymphocytes from a whole-blood sample within 24 hours of patient sampling. A method is described in which blood samples are mixed with a cryopreservative ('Glycigel'), stored frozen, and DNA suitable for use in an HIV PCR recovered. Samples can be stored at -20-degrees-C for up to 3 months and still give positive results with all samples from infected patients; storage at -80-degrees-C for at least 3 months shows no loss of titre. The method shows no loss of sensitivity compared to previously described sample preparation methods. Deglycerolised Glycigel supernatants were found to be suitable for conventional anti-HIV-1 serological studies and loss of sensitivity only represented the dilution effect due to sample preparation. Application of the method as a means of storing samples frozen at the point of sampling and transporting them to a central laboratory for processing is demonstrated using samples taken from HIV-1-infected mothers and their babies.
引用
收藏
页码:217 / 226
页数:10
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