A Collection Of Hiv Rna Nuclisens Easyq Method Evaluation And Its Application Research

BackgroundHIV/AIDS epidemic situation in China is still serious and spreading from high-risk groups to the general population, sexual becomes the main approach of HIV transmission already. Globally, the first important task in HIV prevention and control is to detect HIV early infection, and take measures to prevent the transmission.As it can be detected in peripheral blood after HIV infection, HIV RNA is a crucial auxiliary index for HIV early detection. HIV RNA emerges a high level in acute infections, so it can still be detected when samples are diluted in large amounts of collections. The method of collecting samples to one sample can cut down costs of testing, and some countries have already utilized this pooled test of nucleic acid for routine screening.Currently COBAS AMPLICOR HIV-1 MONITOR Test and Nuclisens HIV-1 are two of common methods for HIV viral load testing in China. A three-stages method of COBAS test with 50 samples was established by the National AIDS Reference Laboratory (NARL), and was used in testing window period among HIV high-risk groups. In comparison to COBAS, EasyQ is a new generation of quantitative method for HIV-1 detection with advantages such as convenience, rapidness, cheaper costs, etc. EasyQ have been now established in most HIV laboratories in domestic region, and the pooled test of which will have great significance in HIV early infections, etc.ObjectivesTo evaluate HIV RNA Nuclisens EasyQ in detecting HIV window period. To assess consistency and correlation of EasyQ and COBAS in testing HIV RNA. To assess application and feasibility of HIV RNA Nuclisens EasyQ in HIV high-risk groups. To provide bases for using HIV RNA Nuclisens EasyQ in practice.Methods1. According to collection procedures, positive RNA samples with 3 levels of virus load copies (one thousand, ten thousands and hundred thousands) were collected in 49 plasma samples which HIV RNA and antibody were negative. After a primary collection was formed, an assessment of repeatability of HIV RNA Nuclisens EasyQ was taken. 2. According to collection procedures, valued serum and quality control plasma samples with 3 levels of virus copies (one thousand, ten thousand and hundred thousand) were collected in 49 plasma samples which HIV RNA and antibody were negative. After a primary collection was formed, a comparison of results of EasyQ and COBAS was carried out by standard operation procedures (SOP) and super sensitive operation procedures (SOP).3. Ten thousand nine hundred and seventy-seven samples from Dehong and Youan hospital were collected and separated, and tested by Nuclisens EasyQ HIV-1 in process of SOP. 2426 samples from injection drug users (IDUs) and spouses of HIV infected in Dehong, and samples from men who have sex with men (MSM) in Youan hospital were tested by the process of collection and separation.4. The experimental data was analyzed using SAS software and Bland-Altman model.Results1. In assay of repeatability of pooled HIV RNA Nuclisens EasyQ, the results of detection in primary collection samples with virus copies of thousand, ten thousand and hundred thousand levels were all positive. One-way analysis of variance showed that repeatability of method was excellent (P=0.9634>0.05).2. Fifty-four primary collection of quality control of goods which were tested by EasyQ and COBAS showed all positive results for HIV-1 RNA, and the correlation between two methods were significance(R=0.855, P<0.0001). The outcomes of consistency showed that mean standard deviation (SD) of two methods was 0.038. Mean±2SD as limit of detection (LOD) for consistency, and there were 51 samples's results in this range of LOD. Finally, we conducted that results of consistency of EasyQ and COBAS were 94.44%(51/54). Paired t-test was conducted to test EasyQ and COBAS, and outcomes showed that two methods had no significance(P>0.05).3. In 54 collections of quality control of goods, the amounts of original samples with 3 levels of virus load (one thousand, ten thousand and hundred thousand levels) were 18. Measurement of consistency and correlation was conducted between EasyQ and COBAS, Results in collection samples with three levels showed that significant correlation (R=0.832,0.873 and 0.866, respectively, P<0.0001)and excellent consistency(94.44%).4. No HIV infected during window period was detected among samples from addiction treatment centre, country marriager and IDUs. And 2 window periods were detected among MSM samples, and 1 window period was detected among spouses of HIV infected samples.1 window period was detected among VCT samples, and was followed until HIV-1 antibody was seroconverted.5. Both methods found 3 samples during window period among 2426 samples. The primary, secondary and original samples of these 3 samples were all positive confirmed by EasyQ and COBAS, others were negative. Outcomes showed that two methods were highly consistency.Conclusions1. This study establishes a test of pooled HIV RNA Nuclisens EasyQ, experimental outcomes show that HIV RNA Nuclisens EasyQ could be feasible and reliable to test collection samples with 3 phases(1:1,1:10,1:50), and suitable for manual operation.2. EasyQ and COBAS have good consistency and correlation to test HIV RNA in collection samples. EasyQ could be feasible to test HIV RNA in collection samples and detect window period, etc.3. Positive result could still be detected in original samples when HIV RNA result is 32IU/ml in primary collection samples, so the pooled HIV RNA Nuclisens EasyQ has an excellent sensitivity.