来自编辑

E. Stoddart
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摘要

编辑:在一项彻底的调查中,Haga等人无法证明炎症性肠病患者肠道标本中存在麻疹病毒RNA (Gut 1996;38: 211 - 5)。尽管他们开发了一种非常灵敏的技术,但他们的方法仍然存在根本性的缺陷。他们描述了肠组织术后切除时间从20到90分钟不等。这种延长的缺血时间会大大降低他们测定的灵敏度,因为会发生大量的RNA降解。MacPherson等人报道,在缺血时间为45分钟至1小时45分钟的手术肠道标本中,RNA的产量明显低于在15秒内冷冻的活检标本。在-70°C的长期储存中也会发生进一步的降解。该试验可能足够敏感,可以检测到一个病毒基因组,但低拷贝数的RNA物种很可能在此过程中丢失。这可以通过试图从提取的肠道RNA样本中扩增一种拷贝数比1-肌动蛋白低得多的肠道RNA来测试。这组作者必须将他们的技术应用于刚切除的肠道组织,才能得出巢式RT-PCR不能检测麻疹、腮腺炎或风疹病毒基因组的结论。史密斯消化内科,北斯塔福德郡医院,纽卡斯尔路,斯托克河畔特伦特,斯塔福德郡ST4 6QG
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From the Editor
EDITOR,-In a thorough investigation, Haga et al were unable to demonstrate the presence of measles virus RNA in intestinal specimens from patients with inflammatory bowel disease (Gut 1996; 38: 211-5). Although they developed an exquisitely sensitive technique, there remains a fundamental flaw in their methodology. They described intestinal tissue postoperative resection times ranging from 20 to 90 minutes. Such prolonged ischaemic times would have reduced the sensitivity of their assay considerably, as substantial RNA degradation would have occurred. MacPherson et al reported a significantly lower yield of RNA in surgical intestinal specimens with ischaemic times of45 minutes to 1 hour 45 minutes compared with biopsy specimens frozen within 15 seconds.' Further degradation can also occur during prolonged storage at -70°C. The assay may be sufficiently sensitive to detect one viral genome, but low copy number RNA species are likely to have been lost on the way. This could be tested by attempting to amplify an intestinal RNA species present in much lower copy numbers than 1-actin from their extracted intestinal RNA samples. The authors must apply their technique to freshly resected intestinal tissue before they can conclude that nested RT-PCR fails to detect measles, mumps or rubella viral genomes. M S H SMITH Department of Gastroenterology, North Staffordshire Hospital, Newcastle Road, Stoke on Trent, Staffordshire ST4 6QG
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