Factors Affecting Isolation and Propagation of Bovine Coronavirus in Human Rectal Tumor-18 Cell Line

Coronavirus infection in intensively managed cattle with respiratory disease

“…BCoV was isolated using human rectal tumour (HRT‐18) cells . Viral transport medium from swabs, or a 20% suspension of tissues homogenised in PBGS, was centrifuged at 560 g for 15 min.…”

Neonatal Enterocolitis Associated with Coronavirus Infection in a Foal: A Case Report

“…The amino acid sequence identities of open reading frames (ORFs) 5, 6, and 7 for US isolate VR-2332 and the European isolate Lelystad virus (LV) are only 59%, 78%, and 65% homologous, respectively. 6,7 In recent years, PRRS has occurred in some regions of P. R. China and has caused severe economic losses in the swine industry. Here, we describe the first isolation of a PRRSV isolate from a pig herd in the central-eastern part of P. R. China.…”

“…22 The primary advantage of the immunohistochemistry test is that it has better sensitivity and specificity than other detection methods. 3,5,7,22 This testing procedure was selected because of the previously identified specificity that has been verified by a lack of monoclonal reactivity with other viral pathogens such as transmissible gastroenteritis virus, feline infectious peritonitis virus, infectious bovine rhinotracheitis virus, bovine viral diarrhea virus, parainfluenza virus, and bovine respiratory syncytial virus by indirect fluorescent antibody test or immunoperoxidase testing procedures. 22 This technique identified coronaviral antigen not only in the cytoplasm but also in the cell nuclei.…”

Comparison of Immunohistochemistry, Electron Microscopy, and Direct Fluorescent Antibody Test for the Detection of Bovine Coronavirus

“…33,34 Although virus isolation is a sensitive method for the detection of animal viruses, no cell culture system is currently available in which all field strains of BCV can grow consistently to a significantly high titer during primary isolation. 21 In addition, virus isolation is time consuming and labor intensive. 37 Immunofluorescence staining of frozen tissue sections is rapid and simple as compared with virus isolation and TEM 29 and is commonly used for the detection of many viruses of veterinary interest.…”

“…35 BCV can be detected by transmission electron microscopic examination of fecal samples, direct fluorescent antibody assay (DFA) on frozen sections of spiral colon, 23 and enzyme-linked immunosorbent assay (ELISA) 10 and occasionally through virus isolation in vitro. 21 Currently, transmission electron microscopy (TEM) and DFA are the most widely used methods for BCV diagnosis. 27 Results from TEM can be obtained in a few hours, but the technique has the limi- tation of detecting nonviral particles that may look like coronavirus and detecting virus only when the number of virions is Ͼ10 6 /g of feces.…”