Development and validation of multiplex one-step qPCR/RT-qPCR assays for simultaneous detection of SARS-CoV-2 and pathogens associated with feline respiratory disease complex

被引:2
|
作者
Thieulent, Come J. [1 ,2 ]
Carossino, Mariano [1 ,2 ]
Peak, Laura [1 ]
Wolfson, Wendy [3 ]
Balasuriya, Udeni B. R. [1 ,2 ]
机构
[1] Louisiana State Univ, Sch Vet Med, Louisiana Anim Dis Diag Lab, Baton Rouge, LA 70803 USA
[2] Louisiana State Univ, Sch Vet Med, Dept Pathobiol Sci, Baton Rouge, LA 70803 USA
[3] Louisiana State Univ, Sch Vet Med, Dept Vet Clin Sci, Baton Rouge, LA USA
来源
PLOS ONE | 2024年 / 19卷 / 03期
关键词
POLYMERASE-CHAIN-REACTION; EQUINE-ARTERITIS-VIRUS; AVIAN INFLUENZA H5N1; TRACT DISEASE; BORDETELLA-BRONCHISEPTICA; DOMESTIC CATS; CHLAMYDIA-FELIS; H3N2; VIRUS; PCR ASSAY; PREVALENCE;
D O I
10.1371/journal.pone.0297796
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Feline respiratory disease complex (FRDC) is caused by a wide range of viral and bacterial pathogens. Both Influenza A virus (IAV) and Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) also induce respiratory diseases in cats. Two one-step multiplex qPCR/RT-qPCR assays were developed and validated: FRA_1 (Feline respiratory assay 1) for the detection of four viral targets and FRA_2 for the detection of three bacteria associated with FRDC. Both multiplex assays demonstrated high specificity, efficiency (93.51%-107.8%), linearity (> 0.998), analytical sensitivity (<= 15 genome copies/mu l), repeatability (coefficient of variation [CV] < 5%), and reproducibility (CV < 6%). Among the 63 clinical specimens collected from FRDC-suspected cats, 92.1% were positive for at least one pathogen and co-infection was detected in 57.1% of samples. Mycoplasma felis (61.9%) was the most found pathogen, followed by feline herpesvirus-1 (30.2%), Chlamydia felis (28.7%) and feline calicivirus (27.0%). SARS-CoV-2 was detected in two specimens. In summary, this new panel of qPCR/RT-qPCR assays constitutes a useful and reliable tool for the rapid detection of SARS-CoV-2 and viral and bacterial pathogens associated with FRDC in cats.
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页数:18
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