ABSTRACT
In this study a nested-PCR assay was optimized for detection of two BVDVbiotype of NADL strain. Apart of 5' non-coding region of virus, 249 bp in size, was amplified in RT-PCR. PCR product was cloned in a pTZ57R/T vector and sequencing results confirmed the specificity of the test. Internal primers were designed and a 155 bp DNAfragment was amplified in nested-PCR. The 4 sensitivity of RT-PCR and nested-PCR for detection of virus in cell culture were found to be 10 2 TCID50 and 10 TCID50, respectively. Seven cell cultures were tested for BVDVcontamination using ELISA, RT-PCR and nested-PCR. Results indicate that sensitivity of molecular tests for detection of virus in cell culture samples is higher than ELISA
Subject(s)
Cells, Cultured , Polymerase Chain Reaction , Enzyme-Linked Immunosorbent AssayABSTRACT
McArthur-Bates Communicative Development Inventory [CDI] is a precise method to examine language in the early years of life. McArthur-Bates CDI was selected to help to improve Persian Language pathology movement to do researches on early intervention for language disorders in early childhood. The purpose of this study was to define the facial and contextual validity and reliability of the translated infant form of CDI in Persian. This study had an analytical-descriptive and cross-sectional design. The sample included thirty 8-16 months old children, without any mental and genetic problems. They were selected non-randomly from the health centers in Isfahan. Parents of selected children completed two questionnaires including basic information and CDI infant form. The answers were analyzed by calculating internal correlation in questions to define facial and contextual validity and by Cronbach alpha criterion to fix reliability coefficient. Face and content validity were determined through considering a linguist's comments as well as two speech therapists and parents' viewpoints about CDI items. The reliability coefficients for different sections of CDI-infant form were computed respectively from the lowest [?lpha=0.43] to the highest [?lpha=0.98] in regard of general gestures and vocabulary comprehension. The results were expectable considering the goal of determining the validity of translation and sample size. However, it seems that item counts of vocabulary comprehension subscale caused its highest reliability and the least cultural compatibility of general gestures with Iran community caused its lowest reliability
ABSTRACT
The aim of the present study was using RT-PCR for the diagnosis of avian infectious bronchitis virus and Massachusetts serotype in tissue samples. Optimization of a molecular diagnostic method for the detection of avian bronchitis virus and identification of Massachusetts serotype was investigated. In order to detect infectious bronchitis virus [IBV] in tissue samples, an RT-PCR was optimized. Specific primers from conserved region of all known IBVserotypes were used in the first PCR assay. Specific primers for the identification of Massachusetts serotype were selected from S-1 gene of the virus in the second PCR reaction. The S-1 gene is a hypervariable region among IBV serotypes; therefore, the amplification of this region is important for serotype identification. Viral RNAwas extracted from vaccine and tissue samples from vaccinated and clinical tissue samples of suspected birds. After construction of cDNA, two PCR assays were performed. In the first RT-PCR, detection of virus in test samples was investigated and in the second PCR, Massachusetts serotype was identified. To identify the specificity of the test, PCR amplicons were sequenced. In the first reaction, the IBVwas detected in the samples and a 600 bp fragment was amplified. In the second PCR, a 355 bp fragment was amplified from S-1 gene, which confirmed the detection of Massachusetts serotype. In clinical samples, the IBVwas also detected. The specificity of the test was confirmed by sequencing of PCR products. Sequence data were submitted to the GenBank which could be accessible by AY954694 number. The RT-PCR is a specific assay for the detection of IBV. Detection of IBV and identification of genetic differences among IBV subtypes in Massachusetts serotype would be possible by sequencing of amplified products