Eight strains of infectious bronchitis virus (IBV) were isolated between 1986 and 1995 from broilers and layers at eight different farms in four provinces in China. The viruses were isolated from flocks which suffered from either respiratory disease or nephritis and the majority had not been vaccinated against IBV. Six strains were shown by monoclonal antibodies to differ from H120, Connecticut and Arkansas 99 strains of IBV and also to differ from each other. Four of these strains were serotyped; one (NRZ) was of the Massachusetts serotype, three (HV, NB-90 and TJ) shared a degree of antigenic similarity and were of a serotype that differed from Massachusetts and Connecticut. NB-90 was similar to both Gray and T strains whereas TJ shared some similarity with the T strain. Four strains, HV, NB-90, YY and TJ induced 33, 47, 60 and 90% mortality, respectively, in 3-week-old specific pathogen-free chickens. Clinical signs and post-mortem findings were identical to those induced by the nephropathogenic T strain. Chickens vaccinated with H120 strain, and then challenged with four highly pathogenic strains HV, NB-90, YY and TJ were not protected as determined by both virus isolation and mortality. The results show that highly pathogenic IBV strains which induce clinical nephritis occur frequently in poultry flocks in China. They also confirm field observations on the lack of protection by currently used IB vaccines of the Massachusetts serotype against challenge with these nephropathogenic strains.

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a b s t r a c tInfectious bronchitis (IB) is a highly infectious viral disease responsible for major economic losses in the poultry industry. A reverse genetic vaccine is a safe, rapid, and effective method of achieving IB prevention and control. In this study, we constructed the recombinant strain, rH120-S1/YZ, using a reverse genetic system, based on the backbone of the H120 vaccine strain, with the S1 gene replaced with that of the QX-like nephropathogenic strain, ck/CH/IBYZ/2011, isolated in China. The results of dwarf chicken embryos, growth kinetics, and viral titration in the embryos demonstrated that the biological characteristics of the recombinant virus remained unchanged. Like the rH120-infected group and in contrast to the rIBYZ-infected group, no mortality, clinical signs, or lesions were observed in the lungs or kidneys of young chickens inoculated with rH120-S1/YZ. The viral loads in various tissues, cloacal, and oral swabs was lower in most types of samples, indicating that the rH120-S1/YZ strain was highly safe in chicks. Compared to rH120 vaccination group, when the efficacy of this strain was evaluated against the QXlike IBV strain, better protection, with 100% survival rate and no disease symptom or gross lesion was observed in the chickens vaccinated with rH120-S1/YZ. Increased levels of IBV-specific antibodies were detected in the serum of the rH120-S1/YZ-vaccinated animals 14 days post-vaccination. Collectively, our results suggest that the recombinant strain, rH120-S1/YZ, may represent a promising vaccine candidate against QX-like IBVs.