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題 名 | Temperature Effect on the Sensitivity of ELISA, PA and WB to Detect Anti-HIV-1 Antibody and Infectivity of HIV-1=溫度效應對粒子凝集法、酵素免疫分析法及西方墨點法檢測抗HIV-1抗體與HIV-1病毒感染力之影響 |
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作 者 | 王躬仁; 楊志元; 林翠莉; 陳豪勇; 洪其璧; | 書刊名 | 中華醫學雜誌 |
卷 期 | 59:6 1997.06[民86.06] |
頁 次 | 頁325-333 |
分類號 | 414.82 |
關鍵詞 | 後天免疫不全症候群; 酵素免疫分析法; 人類免疫不全病毒; 粒子凝集法; 西方墨點法; Acquired immunodeficiency syndrome; Enzyme-linked immunosorbentassay; Human immunodeficiency virus; Particle agglutination; Western blotting; |
語 文 | 英文(English) |
中文摘要 | 背景:後天免疫不全症候群(AIDS)主要是經由性接觸而散佈的一傳染性疾病,截至目前為止仍無有效的療法可以對抗它,因此正確的檢驗結果便顯得意義重大。其中檢體保存的適當與否,被認為可能對檢測結果有所影響。因此本實驗主旨是要評估在溫度較高的情況下及冷凍╱解凍的次數過多時,是否會影響檢驗結果。 方法:測試用血清,在經過不同溫度、時間及冷凍╱解凍的處理後,分別以粒子凝集法(PA)、三種不同廠牌的酵素免疫分析法(ELISA)及西方墨點法(WB)法來檢測anti-HIV抗體是否有任何之改變。而HIV之感染力則是利用syncytia formation assay及聚合酉每連鎖反應(PCR)來測定。 結果:測試用血清,在經過25℃一小時、二小時、四小時;37℃30分鐘、60分鐘;56℃30分鐘、60分鐘;65℃30分鐘及冷凍╱解凍循環七次處理後,對於PA,ELISA,與WB之檢測結果皆沒有任何之影響。只有在65℃下處理一小時,對其中部份ELISA結果產生些微變化,對PA及WB之結果都沒有太大之作用。另外當以ELISA及PA檢測被加熱過之正常血清(50支檢體),並無偽陽性的結果產生。用病毒在感染T細胞後可產生細胞病變(CPE)之syncytia formation assay來檢測HIV之感染力,發現56℃30分鐘、60分鐘及65℃30分鐘、60分鐘的確可消除CPE之產生,但以PCR檢測這些檢體時,卻都可測得HIV-1之DNA,這個數據指出在56℃及65℃之處置下,對檢測結果沒有產生影響,可減少降低HIV之感染力,但無法完全消滅HIV。 結論:在這些處置下,對檢驗結果沒有產生重大之影響。因此往後接anti-HIV抗體測試評估之單位不應該再抱怨因檢體保存不當對其結果會產生重大之改變。另外我們發現先行加熱處理檢體,可以降低HIV之感染力,此舉雖無法完全將HIV去活化,但將可降低暴露在HIV感染之威脅下。然而,必要之防護措施仍須遵守,對保障檢驗操作者之安全,將更有所助益。 |
英文摘要 | Background: This study is designed to resolve the problem of whether temperature or freeze/thaw cycle will have any impact on the sensitivity for detection of anti-HIV-1 antibody by particle agglutination (PA), enzyme-linked immunosorbent assay (ELISA) and western blotting (WB). To reduce potential risk for laboratory personnel exposed to HIV-infection, it will be useful to determine the temperature effect on HIV infectivity. Methods: Testing sera were incubated at different temperatures or treated with several cycles of freeze and thaw. PA, ELISA and WB were used to detect anti-HIV-antibodies, whereas syncytia formation assay and polymerase chain reaction (PCR) were applied to detect HIV-infection. Results: The data showed that certain temperature points (no treatment, 25OC for 1hr, 2hrs and 4 hrs, 37OC for 30 minutes and 60 minutes, 56OC for 30 minutes and 60 minutes, 65OC for 15 minutes and 30 minutes) had no impact on the testing results of ELISA, PA and WB in detection of anti-HIV-1 antibody. In addition, testing results of 50 normal human serum samples which had been heated to 56OC for 30 minutes were still negative by ELISA and PA. Only the samples incubated at 65OC for 60 minutes had slight differences in results. Freeze and thaw treatments of the serum did not alter anti-HIV testing results, either. Treatments of supernatant of HTLV-IIIB culture at 56OC for 30 minutes and 60 minutes, 65OC for 15 minutes and 30 minutes could eliminate the syncytia formation caused by HIV-infection. Further analysis of the samples by PCR was able to detect HIV-specific sequences in all the treatments. Conclusions: Anti-HIV antibody is quite stable in serum, even when it is pre-heated to 56OC for 30 minutes. Freeze and thaw treatment of serum samples up to seven cycles did not change the results, either. In addition, to minimize the potential risk of laboratory personnel exposed to HIV infection, pre-treatment of serum samples with heat at 56OC for 30 minutes or 60 minutes can reduce HIV infectivity. However, laboratories still must emphasize the importance of universal precautions rather than heat-inactivation of serum to prevent occupational transmission of HIV. |
本系統中英文摘要資訊取自各篇刊載內容。