Use of participant EQA results to assess sample homogeneity and stability for qualitative analytes

International requirements for PT and EQA state that providers must demonstrate that samples are homogeneous and stable. However, testing for homogeneity and stability can be expensive, use samples that could otherwise serve as quality-control materials, and can also fail to detect significant inhomogeneity and instability. In some situations it may be preferable to use the results from participants to identify problems with sample lots, if historic results follow predictable distributions and a statistical decision rule can be produced. An unusually high proportion of incorrect results may indicate that samples were inhomogeneous or unstable. Conditions under which this can be demonstrated are discussed, and the efficacy of the proposal is demonstrated with two examples. This procedure is especially effective when there are a large number of participants and/or a historic small proportion of incorrect results. Providers who adopt this proposal will need to retain samples for testing and assume the risk of distributing bad samples.Presented at the Eurachem PT Workshop September 2005, Portorož, Slovenia     

A data-driven,high-throughput methodology to determine tissue-specific differentially methylated regions able to discriminate body fluids

Tissue-specific differentially methylated regions (tDMRs) are regions of the genome with methylation patterns that modulate gene expression in those tissue types. The detection of tDMRs in forensic evidence can permit the identification of body fluids at trace levels. In this report, we have performed a bioinformatic analysis of an existing array dataset to determine if new tDMRs could be identified for use in body fluid identification from forensic evidence. Once these sites were identified, primers were designed and bisulfite modification was performed. The relative methylation level for each body fluid at a given locus was then determined using qPCR with high-resolution melt analysis (HRM). After screening 127 tDMR's in multiple body fluids, we were able to identify four new markers able to discriminate blood (2 markers), vaginal epithelia (1 marker) and buccal cells (1 marker). One marker for each target body fluid was also tested with pyrosequencing showing results consistent with those obtained by HRM. This work successfully demonstrates the ability of in silico analysis to develop a novel set of tDMRs capable of being differentiated by real time PCR/HRM. The method can rapidly determine the body fluids left at crime scenes, assisting the triers of fact in forensic casework.     

杀虾微杆菌血清同源性及血清学检验

以分离于日本对虾（Penaeus japonicus）虾苗的杀虾微杆菌（Microbacterium shrimpcida sp．nov．）代表菌株（HID10406-1）,分别制备全菌（OK）及热处理（121℃作用1h）的菌体（O）免疫原,强化免疫家兔制备相应抗血清,对供试6株杀虾微杆菌纯培养物进行了血清型检定。结果表明,均存在同种的O抗原（血清同源）,不存在不耐热表面（K）抗原。以相应OK抗血清为第一抗体,以标准的羊抗兔IgG荧光抗体为第二抗体,对该6株纯培养菌及用代表菌株人工感染病死虾体组织进行间接荧光抗体染色,结果显示了特异的荧光反应;同时,对从人工感染死亡虾分离的10株纯培养菌以凝集反应方法进行了检验,显示出特异凝集现象。     

Comprehensive quality control programme for serology and nucleic acid testing using an Internet-based application

The National Serology Reference Laboratory, Australia (NRL) has quality assured the serology for high risk blood-borne infections since 1985, commencing with anti-Human Immunodeficiency Virus (HIV) enzyme immunoassays and later extending the programmes to other blood-borne virus testing and to nucleic acid testing. A quality control (QC) programme was considered the most appropriate manner in which to achieve real-time monitoring. An Internet-based application, EDCNet, facilitates the entry of results of QC sample testing and returns the analysed results instantaneously. Results can be displayed in a variety of tables and charts, so that QC results from blood service and diagnostic laboratories can be monitored. Comparison of results between laboratories using the same system offers monitoring of accuracy, while within-laboratory comparisons offer monitoring of the assay precision. More than 200,000 data points were submitted to EDCNet in 2002 from blood service laboratories as well as from diagnostic laboratories. Analysis of reagent batch variability was determined, e.g. the coefficient of variation between batches of seven assays used to detect anti-hepatitis C virus (HCV) antibodies ranged from 4.9% to 21.6%. Using EDCNet, laboratories can monitor both precision and accuracy of results by comparison with the results of other laboratories. The system may be a highly cost-effective method for maintaining quality.