中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2011年
5期
459-464
,共6页
肝炎病毒,乙型%DNA,病毒%试剂盒,诊断%病毒载量%聚合酶链反应%敏感性与特异性
肝炎病毒,乙型%DNA,病毒%試劑盒,診斷%病毒載量%聚閤酶鏈反應%敏感性與特異性
간염병독,을형%DNA,병독%시제합,진단%병독재량%취합매련반응%민감성여특이성
Hepatitis B virus%DNA,viral%Reagent kits,diagnostic%Viral loal%Polymerase chain reaction%Sensitivity and specificity
目的 分析两种试剂在HBV DNA定量检测中的相关性,评价其在不同病毒载量时的检测性能.方法 用人AB型血清将WHO第2代HBV DNA国际标准品(编号:97/750)配制成不同浓度的10份样本,10份样本的浓度分别为1×106、5×106、1×105、5×104、1×104、5×103、1×103、5×102、1×102、1×101 kIU/L.采用深圳匹基生物工程股份有限公司生产的HBV DNA荧光定量PCR检测试剂(PG试剂)和美国罗氏公司生产的定量PCR试剂(罗氏试剂)检测78份HBV感染者血清、30份健康献血者血清和10份不同浓度范围的WHO标准品中HBV DNA含量.对两种试剂检测HBV DNA结果相关性进行分析;对试剂在不同病毒载量时的检测性能进行评价;并对漏检情况进行分析.两种试剂每批检测均设有阴性质控、弱阳性质控和强阳性质控.结果 两种试剂对WHO HBV DNA标准物质稀释血清均能正确检出,罗氏试剂能检出最高稀释度样本浓度为2.00(kIU/L,lg),PG试剂能检出最高稀释度样本浓度为3.00(kIU/L,lg);两种试剂检测结果存在线性相关(R2=0.938 7,P<0.01),罗氏试剂检测上限与理论值相符,大于检测上限的标本经稀释重复测定,罗氏试剂检测结果[(8.35±0.20)kIU/L,lg]高于PG试剂检测结果[(7.73±0.42)klU/L,lg],差异有统计学意义(t=3.776,P<0.05).两种试剂检测108份血清标本中HBV DNA含量,罗氏试剂检测值[(5.88±1.64)kIU/L,lg]高于PG试剂检测值[(5.25±1.55)kIU/L,lg],差异有统计学意义(t=12.297,P<0.01);检测高HBV病毒载量[(>5.00～≤7.00)kIU/L,lg和(>7.00～≤9.00)kIU/L,lg]组,两种试剂的相关性较高(R2分别为0.779 7、0.603 7,P均<0.01);对低HBV病毒载量(>3.00～≤5.00 kIU/L,lg)组,两种试剂的相关性较低(R2=0.417 3,P<0.01);病毒载量为>3.00～≤4.00 kIU/L,lg组,PG试剂的漏检率为33.3%(5/15);病毒载量为>1.08～≤3.00 kIU/L,lg组,PG试剂未检出.结论 PG试剂与岁氏试剂检测结果虽存在一定差距,但相关性较好.两种试剂低病毒载量标本的相关性要低于高病毒载量标本,PG试剂检测HBV DNA的线性范围较窄.
目的 分析兩種試劑在HBV DNA定量檢測中的相關性,評價其在不同病毒載量時的檢測性能.方法 用人AB型血清將WHO第2代HBV DNA國際標準品(編號:97/750)配製成不同濃度的10份樣本,10份樣本的濃度分彆為1×106、5×106、1×105、5×104、1×104、5×103、1×103、5×102、1×102、1×101 kIU/L.採用深圳匹基生物工程股份有限公司生產的HBV DNA熒光定量PCR檢測試劑(PG試劑)和美國囉氏公司生產的定量PCR試劑(囉氏試劑)檢測78份HBV感染者血清、30份健康獻血者血清和10份不同濃度範圍的WHO標準品中HBV DNA含量.對兩種試劑檢測HBV DNA結果相關性進行分析;對試劑在不同病毒載量時的檢測性能進行評價;併對漏檢情況進行分析.兩種試劑每批檢測均設有陰性質控、弱暘性質控和彊暘性質控.結果 兩種試劑對WHO HBV DNA標準物質稀釋血清均能正確檢齣,囉氏試劑能檢齣最高稀釋度樣本濃度為2.00(kIU/L,lg),PG試劑能檢齣最高稀釋度樣本濃度為3.00(kIU/L,lg);兩種試劑檢測結果存在線性相關(R2=0.938 7,P<0.01),囉氏試劑檢測上限與理論值相符,大于檢測上限的標本經稀釋重複測定,囉氏試劑檢測結果[(8.35±0.20)kIU/L,lg]高于PG試劑檢測結果[(7.73±0.42)klU/L,lg],差異有統計學意義(t=3.776,P<0.05).兩種試劑檢測108份血清標本中HBV DNA含量,囉氏試劑檢測值[(5.88±1.64)kIU/L,lg]高于PG試劑檢測值[(5.25±1.55)kIU/L,lg],差異有統計學意義(t=12.297,P<0.01);檢測高HBV病毒載量[(>5.00～≤7.00)kIU/L,lg和(>7.00～≤9.00)kIU/L,lg]組,兩種試劑的相關性較高(R2分彆為0.779 7、0.603 7,P均<0.01);對低HBV病毒載量(>3.00～≤5.00 kIU/L,lg)組,兩種試劑的相關性較低(R2=0.417 3,P<0.01);病毒載量為>3.00～≤4.00 kIU/L,lg組,PG試劑的漏檢率為33.3%(5/15);病毒載量為>1.08～≤3.00 kIU/L,lg組,PG試劑未檢齣.結論 PG試劑與歲氏試劑檢測結果雖存在一定差距,但相關性較好.兩種試劑低病毒載量標本的相關性要低于高病毒載量標本,PG試劑檢測HBV DNA的線性範圍較窄.
목적 분석량충시제재HBV DNA정량검측중적상관성,평개기재불동병독재량시적검측성능.방법 용인AB형혈청장WHO제2대HBV DNA국제표준품(편호:97/750)배제성불동농도적10빈양본,10빈양본적농도분별위1×106、5×106、1×105、5×104、1×104、5×103、1×103、5×102、1×102、1×101 kIU/L.채용심수필기생물공정고빈유한공사생산적HBV DNA형광정량PCR검측시제(PG시제)화미국라씨공사생산적정량PCR시제(라씨시제)검측78빈HBV감염자혈청、30빈건강헌혈자혈청화10빈불동농도범위적WHO표준품중HBV DNA함량.대량충시제검측HBV DNA결과상관성진행분석;대시제재불동병독재량시적검측성능진행평개;병대루검정황진행분석.량충시제매비검측균설유음성질공、약양성질공화강양성질공.결과 량충시제대WHO HBV DNA표준물질희석혈청균능정학검출,라씨시제능검출최고희석도양본농도위2.00(kIU/L,lg),PG시제능검출최고희석도양본농도위3.00(kIU/L,lg);량충시제검측결과존재선성상관(R2=0.938 7,P<0.01),라씨시제검측상한여이론치상부,대우검측상한적표본경희석중복측정,라씨시제검측결과[(8.35±0.20)kIU/L,lg]고우PG시제검측결과[(7.73±0.42)klU/L,lg],차이유통계학의의(t=3.776,P<0.05).량충시제검측108빈혈청표본중HBV DNA함량,라씨시제검측치[(5.88±1.64)kIU/L,lg]고우PG시제검측치[(5.25±1.55)kIU/L,lg],차이유통계학의의(t=12.297,P<0.01);검측고HBV병독재량[(>5.00～≤7.00)kIU/L,lg화(>7.00～≤9.00)kIU/L,lg]조,량충시제적상관성교고(R2분별위0.779 7、0.603 7,P균<0.01);대저HBV병독재량(>3.00～≤5.00 kIU/L,lg)조,량충시제적상관성교저(R2=0.417 3,P<0.01);병독재량위>3.00～≤4.00 kIU/L,lg조,PG시제적루검솔위33.3%(5/15);병독재량위>1.08～≤3.00 kIU/L,lg조,PG시제미검출.결론 PG시제여세씨시제검측결과수존재일정차거,단상관성교호.량충시제저병독재량표본적상관성요저우고병독재량표본,PG시제검측HBV DNA적선성범위교착.
Objective To evaluate clinical significance of two real-time fluorescence quantitative PCR kits for quantitative detection of HBV DNA and detection performance at different viral load levels.Methods A series of calibrators with different concentrations(1×106,5×105,1×105,5×104,1×104,5×103,1×103,5×102,1×102,1×101 kIU/L) were prepared with AB-type sera using the second generation WHO international standard (NIBSC code:97/750). HBV viral load in the sera of 78 patients,30 healthy blood donors and 10 calibrators were detected by real-time fluorescence quantitative PCR HBV DNA test kit from PIJI Bio-Technical Development Company Ltd (PG kit) and Cobas AmpliPrep/Cobas TaqMan HBV test kit. The correlation of the two methods was evaluated, and the performance of the two kits different viral load levels was evaluated. The false negative rate was analyzed. Negative control, low positive control and high positive control were included in every batch. Results Both two kits showed the correct results for the 10 specimens from the WHO international standards. The lowest detection limit of HBV DNA for Roche Cobas AmpliPrep/Cobas TaqMan HBV test and PG kit were 2.00 (kIU/L, lg) and 3.00 (kIU/L,lg) ,respectively. There was linear correlation between the results from Roche Cobas AmpliPrep/Cobas TaqMan HBV test and PG kit ( R2=0.938 7, P < 0.01 ), the upper limit of Roche kit had coincided with theoretical value. The samples with HBV DNA level above the upper limit of detection were diluted and retested to obtain the precise result. The result form Roche Cobas AmpliPrep/Cabas TaqMan HBV test [(8.35±0.20) kIU/L, lg] was higher than that from PG kit [(7.73±0.42 ) kIU/L, lg] (t=3. 776, P <0.05) . The detection of 108 serum samples showed that the level of HBV DNA detected by Roche Cobas AmpliPrep/Cobas TaqMan HBV test [(5.88±1.64) kIU/L, lg] was higher than that by PG kit [(5.25±1.55 kIU/L,lg] (t=12. 297 ,P <0.01 ). The correlation coefficients were high in samples with high HBV viral load[HBV DNA(>5.00 and≤7.00) kIU/L,Ig,R2=0. 779 7, P <0.01 ;HBV DNA( >7.00 ands≤9.00) kIU/L,lg,R2=0.603 7, P <0.01]. The correlation coefficient was low in samples with low HBV viral load[HBV DNA ( > 3.00 and≤5.00) kIU/L, lg, R2=0. 417 3, P <0.01 )]. When HBV DNA ( >3.00 and≤4.00) kIU/L,lg,the false negative rate was 33.3% (5/15). When HBV DNA ( > 1.08and≤3.00) kIU/L,lg,none of positive samples was detected with PG kit. Conclusions PG kit is not as good as Cobas AmpliPrep/Cobas TaqMan HBV test . The linear correlation between the results from the two kits is good. The correlation between the results detected with PG kit and Cobas AmpliPrep/Cobas TaqMan HBV test is higher in the high viral load groups than in the low viral load group. It is suggested that PG kit had a narrower linear range.
