Implementation of internal laboratory quality control procedures for the monitoring of ELISA performance at a regional veterinary laboratory.

Quality control (QC) procedures for antigen detection enzyme-linked immunosorbent assays (ELISAs) for hog cholera (HC) virus, foot and mouth disease (FMD) virus, and an antibody detection ELISA for FMD virus were established at a regional veterinary laboratory in northern Thailand. A recently developed computer software package, QCEL, was used to facilitate management and analysis of QC data. The program was used to assess test performance by producing Shewhart-CUSUM control charts which monitored control data for unacceptable fluctuations or trends. QCEL-generated control charts and analyses are presented and discussed. The use of a simple integrated computerised system for storage and analysis of QC control data provided the laboratory with the opportunity to achieve increased confidence in the results of tests performed.

Use of combined Shewhart-CUSUM control charts in internal quality control of enzyme-linked immunosorbent assays for the typing of foot and mouth disease virus antigen.

An enzyme-linked immunosorbent assay (ELISA) for the typing of foot and mouth disease virus (FMDV) antigen was employed for the routine laboratory diagnosis of FMD at a regional veterinary laboratory in northern Thailand. An objective procedure was developed to monitor the test performance of the ELISA, using absolute test control limits in a Shewhart-CUSUM (cumulative sum) control chart method. The procedure detected significant data trends and 'beyond control limit' situations for each antigen typing system (types O, A and Asia 1), using an assay variable (gamma i). Retrospective analysis using Shewhart-CUSUM control charts of data from 42 ELISAs demonstrated that control limits were exceeded in two assays for FMDV type A. The Shewhart-CUSUM control chart is a simple and reliable internal quality control method for the detection of significant random and systematic variation in assays.

Establishment of a typing enzyme-linked immunosorbent assay for foot and mouth disease antigen, using reagents against viruses endemic in Thailand.

Antisera were produced at a central laboratory in Thailand against the endemic serotypes (O, A and Asia 1) of foot and mouth disease (FMD) virus. At a regional veterinary laboratory, these antisera were used in an indirect sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and serotyping of FMD virus (FMDV) antigen. ELISA readings of < 0.10 optical density (OD) units were considered negative. This was verified using fifty tissue samples which were known to be negative for FMDV. The highest mean sample value for three different dilutions was 0.02 OD units. Of a total of 93 samples submitted for antigen typing, 80 (86%) tested positive by ELISA and 13 (14%) were negative. No FMDV was detected in ELISA-negative samples following attempted tissue-culture virus isolation.

A comparison of type O foot and mouth disease virus field isolates from northern Thailand.

A survey of type O foot and mouth disease (FMD) virus isolates from northern Thailand was undertaken to determine the relationship between field viruses and the vaccine in use, and to gauge the range of antigenic variation among field viruses. Isolates were collected from the two most recent epizootics, 1986-1987 and 1989-1990, and assessed using a two-dimensional neutralisation test to determine their relationship to FMD type O1 Bangkok 1960 (O BKK/60) reference (vaccine challenge) virus. The critical r value for the survey was 0.259 and all isolates tested were found to have an r value considerably greater than this (range 0.66 to 0.80). The results showed close antigenic relationships between the isolates and the reference virus, and indicated a relatively small range of antigenic variation between the isolates.