Skip to content

Advertisement

  • Methodology
  • Open Access

Development of a sensitive and specific xMAP assay for detection of antibodies against infectious laryngotracheitis and bronchitis viruses

Contributed equally
Virology Journal201815:146

https://doi.org/10.1186/s12985-018-1048-x

  • Received: 19 April 2018
  • Accepted: 20 August 2018
  • Published:

Abstract

Background

A serological method to simultaneously detect antibodies against infectious laryngotracheitis virus (ILTV) and infectious bronchitis virus (IBV) is imperative for the differential diagnosis and evaluation of antibodies titers after vaccination.

Method

The microspheres coated with purified recombinant glycoprotein D (gD) of ILTV or nucleocapsid (N) protein of IBV were incubated with serum samples. The simultaneous quantification of ILTV and IBV antibodies were achieved through the interrogation of microspheres by Luminex 200 detection system. .

Results

This xMAP detection demonstrated no nonspecific reactions with avian influenza virus (AIV), avian leukosis virus (ALV), newcastle disease virus (NDV), and Marek’s disease virus (MDV). The results also demonstrated that the xMAP assay was four times more sensitive than the enzyme-linked immunosorbent assay (ELISA) for ILTV detection and two times more sensitive for IBV detection. A total of 90 chicken serum samples from a chicken farm were tested by xMAP and ELISA assays. The results showed that the coincidence rates were 84.44 and 100% for ILTV and IBV detection, respectively.

Conclusion

This study exhibited an opportunity for the differential diagnosis through simultaneous detection of multiplex antibodies in serum and can be used for the multiplex antibodies evaluation after vaccination.

Keywords

  • Luminex xMAP technology
  • Infectious laryngotracheitis virus
  • Infectious bronchitis virus, antibody detection, ELISA

Background

Infectious laryngotracheitis (ILT) and infectious bronchitis (IB) are common respiratory diseases in poultry and are currently present at epidemic levels around the world, including in China, the United States, and India. Because ILT and IB have high incidence and infectivity in chickens at different ages (in days), they have caused huge economic losses to the poultry industry. Both diseases have similar clinical symptoms and pathological changes, leading to significant difficulties in clinical differential diagnosis, especially in cases of mixed infection [1, 2]. In addition, the use of vaccines is the main approach to control of the economically important poultry viral respiratory diseases, such as infectious laryngotracheitis and infectious bronchitis [3]. Therefore, it is important to establish a method to simultaneously detect ILTV and IBV antibodies for the differential diagnosis and immune response evaluation after vaccination.

Infectious laryngotracheitis virus (ILTV), an alphaherpes virus, possesses at least 10 envelope glycoprotein, two main proteins of which are the glycoprotein B (gB) and gD, respectively, which are highly conserved herpesvirus structural glycoproteins [4]. The gD of herpes virus has an important role by binding to the host receptors [1, 5]. The gD protein has been demonstrated to be a candidate antigen for recombinant vaccines [6, 7]. The infectious bronchitis virus (IBV) genome encodes four major structural proteins: the spike glycoprotein (S), the membrane glycoprotein (M), the nucleocapsid (N) protein, and the envelope or small membrane protein (E) [8]. The N protein is thought to be an appropriate diagnostic reagent for antibody detection [9]. In this study, gD and N proteins were selected as antigen molecules for the diagnosis of ILT and IB.

At present, the methods used for the diagnosis and effect evaluation of vaccine immunity of avian respiratory diseases primarily include virology detection, serological detection, and molecular biological detection. Traditional methods, such as virus isolation, animal inoculation experiments, ELISA, hemagglutination (HA), and hemagglutination inhibition (HI) assays are characterized by complex procedures and long periods of time required for diagnosis. Despite its unique, sensitive, simple features and rapidity, polymerase chain reaction (PCR) is unable to meet the requirements for high-flux quarantine and is not favorable for the urgent screening of bulk samples. Therefore, it is imperative to establish a new technology for the effect evaluation of vaccine immunity and the rapid differentiation or correct identification of important avian respiratory diseases.

A new high-flux detection technology, flexible xMAP (x = Unknown, MAP = Multi-Analyte Profiling) assay, uses variously colored polystyrene beads that is carboxylated to allow covalent coupling of protein. Conjugated beads can be used to capture specific antibodies in serum, and a fluorescent secondary antibody is incubated to bind to the captured serum antibodies. A red laser is used to determine the color of the bead and a green laser to detect fluorescence intensity of bound secondary antibodies through the Luminex 200 detection system [10]. This method is applicable for the simultaneous and rapid detection of multiplex pathogen,s antibodies, with simple operation and high accuracy that are superior to conventional methods. In this study, a method employing Luminex xMAP technology to simultaneously detect ILTV and IBV antibodies in serum was established, optimized and used for the differential diagnosis of IBV and ILTV. This assay can also be used to simultaneous monitoring of IBV and ILTV antibody levels for the evaluation of immunity after vaccination.

Results

Establishment of singleplex xMAP detection for ILTV and IBV

A singleplex xMAP for ILTV was established by testing six concentrations of the ILTV gD envelope protein (2.5 μg, 5 μg, 10 μg, 20 μg, 40 μg, and 80 μg per 5 × 105 magbeads), seen in Table 1. The P/N value is the ratio of MFI (Median Fluorescence Intensity) value between the positive sample and negative sample. The optimal conjugation protein concentration was 2.5 μg per 5 × 105 magbeads for singleplex xMAP detection of ILTV. The mean P/N value under the condition was 10.18.
Table 1

P/N values for ILTV antibody xMAP detection

Serum dilution

Coating antigen dilutions (μg/5 × 105 beads)

Average

2.5

5

10

20

40

80

1:25

16.84

8.41

11.18

10.15

6.87

3.22

9.44

1:50

10.47

6.26

8.54

7.39

4.43

2.70

6.63

1:100

8.30

5.07

5.84

6.11

4.07

2.69

5.35

1:200

5.14

3.88

4.83

4.55

2.97

2.21

3.93

Average

10.18

5.90

7.60

7.05

4.59

2.70

 
A singleplex xMAP for IBV was established by testing six concentrations of the IBV N envelope protein (3 μg, 6 μg, 12 μg, 24 μg, 48 μg, and 96 μg per 5 × 105 magbeads), seen in Table 2. The P/N value is the ratio of MFI value between the positive sample and negative sample. The optimal conjugation protein concentration was 12 μg per 5 × 105 magbeads for singleplex xMAP detection of IBV. The mean P/N value under the condition was 17.16.
Table 2

P/N values for IBV antibody xMAP detection

Serum dilution

Coating antigen dilutions (μg/5 × 105 beads)

Average

3

6

12

24

48

96

1:50

13.16

14.67

17.53

14.75

16.30

13.30

14.95

1:100

12.23

18.15

16.44

16.07

18.83

17.66

16.57

1:200

13.41

17.07

17.73

14.63

15.16

17.52

15.92

1:400

11.59

14.72

16.93

11.88

12.30

14.90

13.72

Average

12.60

16.15

17.16

14.33

15.65

15.85

 

Establishment of duplex xMAP for simultaneous detection of ILTV and IBV

Based on the results in Tables 1 and 2, the optimum serum dilution ratio for ILTV and IBV were 1:25 and 1:100, respectively. When singleplex xMAP assay was performed. To determine the optimal serum dilition ration for simultaneous detection of ILTV and IBV, a serial serum dilutions from the range of 1:25 to 1:200 were prepared and were incubated with ILTV and IBV antigen conjugated microspheres. From the P/N values showed in Fig. 1, it is obvious that the optimal serum dilution ratio for duplex xMAP assay was 1:100.
Fig. 1
Fig. 1

Determination of optimal serum dilution ratio for duplex xMAP assay for ILTV and IBV antibodies. ILTV- and IBV-positive and negative sera were diluted at the range from 1:25 to 1:200. MFI Values were obtained after the interrogation of microspheres coated with proteins of ILTV or IBV using Luminex 200 detection system. The P/N values were used to determine the optimal dilution ratio at 1:100 for duplex detection of ILTV and IBV antibodies

Determination of threshold

The threshold is considered as the sum of the average plus three times of standard deviation, which is also named cut-off value. To determine the threshold of xMAP assay for ILTV and IBV, 32 special pathogen free chicken serum samples at 1:100 dilution were used to obtain the average MFI value and standard deviation. From the Table 3, it showed that the threshold of ILTV and IBV detection were 154.95 and 266.75, respectively.
Table 3

Threshold of ILTV and IBV antibodies detection

 

ILTV MFI value

IBV MFI value

SPF chicken negative sera

113

101.5

105

93

207.5

198.5

196.5

184

105

97

110

134

192.5

197

222.5

258

68

68

70

74.5

127

132

134.5

149.5

58

67

68

72

116.5

134

135

141.5

106

115.5

75

90

183

152.5

148

134

126

98

104.5

111

140

158.5

164

116

89.5

86

77

86

132.5

150

153

144

48.5

59.5

43

70

119

95

132

87

Average

87.17

154.22

Standard deviation

22.59

37.51

Threshold

154.95

266.75

Specificity of xMAP assay for ILTV and IBV

The results in Figs. 2 and 3 demonstrated that xMAP duplex assay for ILTV and IBV has a high specificity since there were no cross-reactions with serum positive for other avian diseases, such as avian influenza virus (AIV), avian leukosis virus (ALV), newcastle disease virus (NDV), and Marek’s disease virus (MDV).
Fig. 2
Fig. 2

Specificity for duplex xMAP assays for ILTV antibody detection. Positive sera for ILTV, IBV, ALV, NDV, MDV, AIV(H5, H7 and H9) were used to determine the specificity of duplex xMAP assay. The specific detection results showed that there were no cross-reactions with positive sera for other avian diseases

Fig. 3
Fig. 3

Specificity for duplex xMAP assays for IBV antibody detection. Positive sera for ILTV, IBV, ALV, NDV, MDV, AIV were used to determine the specificity of duplex xMAP assay. The specific detection results showed that there were no cross-reactions with positive sera for other avian diseases

Repeatability of xMAP assay for ILTV and IBV

Intra-assay repeatability experiments showed that the coefficient of variation (CV) was 5.09% for ILTV and 8.05% for IBV. Inter-assay repeatability experiments showed that the CV was 8.11% for ILTV and 9.48% for IBV. All CVs were less than 10%, indicating that the method has high repeatability. The results were shown in Tables 4 and 5.
Table 4

Repeatability experiment of ILTV antibody detection

ILTV detection

Samples

MFI

Average

Standard deviation

Intra Cv

Repeat Experiment 1

repeat 1

1145.5

1208.33

61.5433

5.09%

repeat 2

1268.5

repeat 3

1211

Negative

111.33

/

/

/

Repeat Experiment 2

repeat 1

1127.5

1143.00

55.8860

4.89%

repeat 2

1205

repeat 3

1096.5

Negative

84.33

/

/

/

Repeat Experiment 3

repeat 1

1046

1028.00

16.3707

1.59%

repeat 2

1024

repeat 3

1014

Negative

75.33

/

/

/

Inter MFI Average

1126.44

   

Standard deviation

91.30

   

Inter Cv

8.11%

   
Table 5

Repeatability experiment of IBV antibody detection

IBV

Samples

MFI

Average

Standard deviation

Intra Cv

Repeat Experiment 1

repeat 1

2096.5

2177.67

75.2103

3.45%

repeat 2

2245

repeat 3

2191.5

Negative

177

/

/

/

Repeat Experiment 2

repeat 1

1770

1884.50

151.6583

8.05%

repeat 2

2056.5

repeat 3

1827

Negative

141.33

/

/

/

Repeat Experiment 3

repeat 1

1842

1832.00

106.3532

5.81%

repeat 2

1721

repeat 3

1933

Negative

127

/

/

/

Inter MFI Average

1964.72

   

Standard deviation

186.27

   

Inter Cv

9.48%

   

Comparison of xMAP assay with ELISA using clinical samples

The sensitivities of xMAP and ELISA were compared using 1:50 to 1:3200 ILTV-positive sera and 1:100 to 1:6400 IBV-positive sera. The results showed that xMAP detected ILTV-positive sera at 1:1600 and IBV-positive sera at 1:3200, while ELISA detected ILTV-positive sera at 1:400 and IBV-positive sera at 1:1600 (see Table 6).
Table 6

Comparison of xMAP and ELISA sensitivity

ILTV detection

IBV detection

Dilution

ELISA S/P

xMAP MFI

Dilution

ELISA S/P

xMAP MFI

1:50

2.9178

+

2205

+

1:100

1.5430

+

2732.5

+

1:100

2.0395

+

1725.5

+

1:200

1.0445

+

2103

+

1:200

1.3866

+

1235

+

1:400

0.6319

+

1960

+

1:400

0.8119

+

806.5

+

1:800

0.3961

+

1161

+

1:800

0.4607

486.5

+

1:1600

0.2301

+

779.5

+

1:1600

0.2536

277.5

+

1:3200

0.1372

428.5

+

1:3200

0.1262

145

1:6400

0.0828

226

Cut-off

0.5000

 

154.95

 

Cut-off

0.2000

 

266.75

 
To compare duplex xMAP assay with ELISA for ILTV/IBV detection, 90 chicken serum samples from a chicken farm in Huizhou, China were used and the results are shown in Table 7. For ILTV, xMAP detected 76 positive cases whereas ELISA detected 69 positive cases; among them, 67 samples were detected positive by xMAP assay and ELISA. All 90 samples were detected as positive for IBV by both methods.
Table 7

Comparison of xMAP and ELISA for ILTV and IBV antibody detection

 

Detection method

Positive sample

Negative samples

Total coincidence

ILTV

xMAP

76 (67a)

14 (9a)

84.44%

ELISA

69

21

 

IBV

xMAP

90 (90a)

0 (0a)

100.00%

ELISA

90

0

 

aIdentical number of detection results for xMAP and ELISA

Discussion

IB, an acute infectious disease characterized by acute watery diarrhea, kidney necrosis, and high mortality, is caused by IBV. Thus far, IB has resulted in economic losses to the poultry industry in various countries [8, 11]. IBV infects chickens at different ages. Chicks infected with IBV are characterized by noisy breathing, panting, sneezing, nasal fluid, and other symptoms; furthermore, laying hens exhibit significantly reduced egg production, broilers exhibits low growth and bad meat quality, and breeding hens exhibit significantly reduced breeding efficiency [12]. ILT is an acute, contact respiratory infectious disease in chickens. Chickens infected with ILT are characterized by nasal fluid, conjunctivitis, panting, coughing, and the production of bloody mucus, as well as swelling, erosion, necrosis, and extensive bleeding in the throat after analysis; laying hens exhibit reduced egg production or even no production, as well as poor egg quality [13]. The disease spreads rapidly and has a wide distribution. The high mortality rate (> 70%) results in huge economic losses to the poultry industry [4]. Given the similar clinical symptoms between ILT and IB, the development of diagnoses for both diseases is significant.

This is the first case report of differential diagnosis of ILTV and IBV with xMAP assay. Luminex xMAP assay is an emerging technology that uses small carboxylated polystyrene microspheres that are internally dyed with both a red and an infrared fluorophore [14]. By changing the ratio of the two fluorophores, it is possible to distinguish up to 100 different color-coded microsphere sets, and each microsphere set may be coupled with a different biological probe. The microspheres are detected and characterized by a dedicated flow cytometer [15]. The Luminex xMAP technology is useful for many different applications. One review describes the use of this technology for the multiplex detection of viral, bacterial, parasitic, and fungal agents using the microsphere-based multiplex nucleic acid assay (MBMNA) and the microsphere-based multiplex immunoassay (MBMI) [16]. MBMIs are typically biochemical tests that allow the detection or measurement of the concentration of a protein in a solution via an antibody or immunoglobulin [17]. MBMIs are often used in the diagnosis of various pathogens, such as human papillomavirus and human cytomegalovirus, to detect antibodies in serum samples [1820]. The advantages of this technology are its simple operation, high accuracy, and reduction in the number required experiments and samples because a single test simultaneously identifies a variety of pathogens.

This study describes the establishment and validation of single xMAP and duplex xMAP assays. First, to achieve effective and rapid clinical diagnosis of ILT and IBV, single xMAPs were established for the separate diagnosis of these two diseases after screening to identify the best concentrations of microsphere antigens for ILTV gD and IBV N (2.5 μg/5 × 105 magbeads and and 12 μg/5 × 105 magbeads, respectively), with an optimal serum dilution of 1:100. However, it is not ideal to diagnose only one type of pathogen antibody because more than one disease often affects the respiratory tract in poultry. Given the similar clinical symptoms of ILT and IBV, the differential diagnosis of these two pathogens with two single xMAP assays using one sample wastes time and energy. To achieve the rapid differential diagnosis of clinical samples, it is necessary to establish a duplex xMAP. In addition to ILTV and IBV, AIV and NDV are also common respiratory viruses for poultry. These four diseases have high incidences, and it is difficult to make a differential diagnosis based on clinical symptoms, which are all similar. Figures 2 and 3 showed that our xMAP methods was able to simultaneously detect the antibodies to ILTV or IBV in the sera and had the high specificity. A duplex xMAP can quickly identify ILTV and IBV infection, but it does not indicate possible infection by AIV and NDV. Therefore, based on this study, we propose that quadruplex xMAP should be established in the future for the detection of ILTV, IBV, AIV and NDV as a fast and effective differentiation tool for these four viruses.

Since ELISA was established to detect ILTV antibodies in 1982 [21], it has demonstrated strong advantages, as well as higher sensitivity than virus neutralization tests (VNT), making it widely applicable. ELISA antibody detection of IBV is also feasible at earlier time points [22, 23] and is currently broadly applied in the poultry industry. Single-analyte ELISA does not support the detection of multiple specific antibody responses simultaneously for a single serum sample [24] and possesses other disadvantages, such as the requirement for a relatively large amount of sample, negligible nonspecific binding, and increased background. However, with the development of the poultry industry, requirements dictate that many samples must be detected simultaneously; thus, a new detection method for diagnosis with ILTV and IBV is significant. Luminex xMAP assay represents an alternative for commonly employed indirect tests such as ELISA. The conversion of an ELISA to the xMAP format is uncomplicated, efficient and cost-saving, producing an assay with superior dynamic range and sensitivity [25]. MBMI represents increased sensitivity and the potential to quantify antibodies, antigens, and other substances (e.g., hormones, cytokines, and tumor markers), unlike conventional ELISA tests [26]. Luminex xMAP has also been compared with a western blot assay for antibody detection [27]. The results obtained from both methods showed that the sensitivity of xMAP was 82.7%, while the western blot assay sensitivity was 74.7%. Therefore, xMAP may be more efficient and precise than western blots for the diagnosis of diseases. Although ELISA provides relatively accurate results and has been used for many years, this study demonstrates that xMAP is much more sensitive than ELISA for the diagnosis of ILT, as indicated by the results presented in Table 6. In addition, xMAP may be used for the rapid diagnosis of many samples, saving time and effort. The results presented in Table 7, demonstrate that xMAP is highly accurate, with similar detection results to those obtained by ELISA in positive clinical samples; thus, xMAP is able to be widely applicable.

Conclusions

This study describes how to determine the optimal conditions for establishing duplex xMAP assay based on the singleplex xMAP result, thus providing a reference for establishing multiplex xMAP assay for simultaneous detection of animal pathogen,s.antibodies.

Methods

Antigens and sera

The antigens used for ILTV were a recombinant gD [28], expressed, purified, preserved in our laboratory. For the detection of antibodies against IBV, the recombinant N protein was expressed as GST fusion protein in Escherichia coli BL21 (DE3) cells [29]. The negative sera of SPF chickens and the positive virus serum (ILTV, ALV, NDV, MDV, H5, H7 and H9) were purchased form Harbin Weike Biological Technology Development Company, China. The positive sera (IBV and AIV) were obtained from China Veterinary Culture Collection Center. Ninety of clinical chicken serum samples were collected from a chicken farm in Huizhou, China.

Coupling of proteins to fluorescent microspheres

Prior to coupling, the recombinant proteins was desalted by gel filtration using MicroBio-Spin 6 columns (Bio-Rad, California, USA) based on the manufacture’s protocol to exchange the buffer from sodium azide or imidazole to PBS. All the antigens were quantified using Pierce BCA protein Quantification Kit (Thermo Scientific, USA). Coupling of proteins to fluorescent microspheres was performed as described by Karanikola et al. [30]. MAP MC10007 beads (Luminex, USA) were coupled by gD protein of ILTV and MAP MC10015 beads (Luminex, USA) were coupled by N protein of IBV. A certain number of beads was transferred to one coupling reaction tube, followed by 100 μL bead wash buffer and suspended in 80 μL bead activation buffer. Then 10 μL of 50 mg/ml EDAC (Sigma-Aldrich) and 10 μL of 50 mg/ml S-NHS (Sigma-Aldrich, Germany) were added in the reaction buffer. The reaction tube was gently vortexed for 20 min at room temperature (RT). PBS (pH 7.4, 150 μL) was added twice and the recombinant protein was added. Incubation was carried out at RT for 2 h. The coupled beads were centrifuged and washed. Ultimately, the beads were stored at 4 °C in 150 μL storage buffer in the darkness.

Methods for bead-based xMAP technology

The xMAP assays were carried out in 96-well polystyrene microplates using Luminex 200 detection system (Luminex, USA). Relevant positive and negative sera have been prepared for the establishment of the singleplex xMAP assay to detect ILTV antibody or IBV antibody in the serum. ILTV gD protein at the following concentrations (2.5 μg, 5 μg, 10 μg, 20 μg, 40 μg, and 80 μg) and IBV N protein at the following concentrations (3 μg, 6 μg, 12 μg, 24 μg, 48 μg, and 96 μg) were coupled with 5 × 105 magbeads to determine the optimum antigen concentration for singleplex xMAP assay. A serious dilution of sera at the range from 1:25 to 1:200 have been made to determine the optimal sample dilution. Cross-reactivity or specificity was evaluated by making the assay with sera from chicken infected with AIV, ALV, NDV, and MDV. Then, a duplex xMAP assay was established based on the result of two separated singleplex xMAP assay. In briefly, a 50 μL aliquot of the simplex beads (50 beads/μL) or the duplex beads (100 beads/μL, 50 beads/μL each kind) was transferred into the wells, added in 50 μL of diluted sera. Incubation was conducted on a plate shaker (800 rpm) at RT for 60 min. The centrifugation was conducted in the magnetic separator for 60 s. The supernatant was removed from each well. Beads were washed 5 times in 100 μL PBS. Each well was added with 100 μL of a biotinylated secondary antibody (goat anti-chicken IgY, Beijing Solarbio Science & Technology, China) at 2 μg/mL. The plate was shaken (800 rpm) for 30 min at RT and washed as described above. Finally, 2 μg/mL of streptavidin-phycoerythrin (SAPE, Life Technologies GmbH) was added to each well at 100 μL/well. The plate was shaken and the supernatant was removed. Finally, 100 μL of assay buffer was added to each well. The plate was shaken for approximately 15 s and analyzed according to the manufacturer’s protocol. All samples were conducted in triplicates in each assay.

ELISA used for comparison

Commercial ILTV and IBV ELISA kit (Biochek, Netherlands) were used to detect presence of ILTV or IBV antibodies in poultry sera. The assay was performed according to the manufacturer’s protocols.

Notes

Abbreviations

AIV: 

Avian influenza virus

ALV: 

Svian leukosis virus

IB: 

Infectious bronchitis

ILT: 

Infectious laryngotracheitis

MDV: 

Marek’s disease virus

NDV: 

Newcastle disease virus

Declarations

Funding

The study was supported by the Science and Technology Research Program from Department of Science and Technology, Guangdong Province and Guangzhou Municipal (2015B070701003, 2016A040403060, 2017B030314171, 2016201604030059 and 2017A070702001) of the. This study was also supported by the National Key Technology R&D Program (2015BAI07B01).

Availability of data and materials

All data generated or analyzed during this study are included in this published article. Contact corresponding author for requests.

Authors’ contributions

All authors contributed to the interpretation of the data, critically revised the manuscript for important intellectual content, approved the final version to be published, and agree to be accountable for all aspects of the work.

Ethics approval and consent to participate

Not applicable.

Consent for publication

Our manuscript does not contain any individual person’s data in any form.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Authors’ Affiliations

(1)
Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China
(2)
Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510640, China

References

  1. Eisenberg RJ, Atanasiu D, Cairns TM, Gallagher JR, Krummenacher C, Cohen GH. Herpes virus fusion and entry: a story with many characters. Viruses-Basel. 2012;4:800–32.View ArticleGoogle Scholar
  2. Ignjatovic J, Sapats S. Avian infectious bronchitis virus. Revue Scientifique Et Technique De L Office International Des Epizooties. 2000;19:493–508.View ArticleGoogle Scholar
  3. Jones RC. Viral respiratory diseases (ILT, aMPV infections, IB): are they ever under control? Br Poult Sci. 2010;51:1–11.View ArticleGoogle Scholar
  4. Fuchs W, Veits J, Helferich D, Granzow H, Teifke JP, Mettenleiter TC. Molecular biology of avian infectious laryngotracheitis virus. Vet Res. 2007;38:261–79.View ArticleGoogle Scholar
  5. Di Giovine P, Settembre EC, Bhargava AK, Luftig MA, Lou H, Cohen GH, Eisenberg RJ, Krummenacher C, Carfi A. Structure of herpes simplex virus glycoprotein D bound to the human receptor nectin-1. PLoS Pathog. 2011;7:e1002277.View ArticleGoogle Scholar
  6. Lazear E, Whitbeck JC, Ponce-de-Leon M, Cairns TM, Willis SH, Zuo Y, Krummenacher C, Cohen GH, Eisenberg RJ. Antibody-induced conformational changes in herpes simplex virus glycoprotein gD reveal new targets for virus neutralization. J Virol. 2012;86:1563–76.View ArticleGoogle Scholar
  7. Zhao W, Spatz S, Zhang ZY, Wen GY, Garcia M, Zsak L, Yu QZ. Newcastle disease virus (NDV) recombinants expressing infectious Laryngotracheitis virus (ILTV) glycoproteins gB and gD protect chickens against ILTV and NDV challenges. J Virol. 2014;88:8397–406.View ArticleGoogle Scholar
  8. Cavanagh D. Coronavirus avian infectious bronchitis virus. Vet Res. 2007;38:281–97.View ArticleGoogle Scholar
  9. Zhang DY, Zhou JY, Fang J, Hu JQ, Wu JX, Mu AX. An ELISA for antibodies to infectious bronchitis virus based on nucleocapsid protein produced in Escherichia coli. Veterinarni Medicina. 2005;50:336–44.View ArticleGoogle Scholar
  10. Krishhan VV, Khan IH, Luciw PA. Multiplexed microbead immunoassays by flow cytometry for molecular profiling: basic concepts and proteomics applications. Crit Rev Biotechnol. 2009;29:29–43.View ArticleGoogle Scholar
  11. Reagan RL, Yancey F, Brueckner AL. Response of young rabbits to infectious bronchitis virus (Wachtel strain). Poult Sci. 1955;34:1173–4.View ArticleGoogle Scholar
  12. Jackwood MW. Review of infectious bronchitis virus around the world. Avian Dis. 2012;56:634–41.View ArticleGoogle Scholar
  13. Swayne DE. Diseases of poultry. 13th ed; 2017.Google Scholar
  14. Dunbar SA, Vander Zee CA, Oliver KG, Karem KL, Jacobson JW. Quantitative, multiplexed detection of bacterial pathogens: DNA and protein applications of the Luminex LabMAP (TM) system. J Microbiol Methods. 2003;53:245–52.View ArticleGoogle Scholar
  15. Earley MC, Vogt RF, Shapiro HM, Mandy FF, Kellar KL, Bellisario R, Pass KA, Marti GE, Stewart CC, Hannon WH. Report from a workshop on multianalyte microsphere assays. Cytometry. 2002;50:239–42.View ArticleGoogle Scholar
  16. Reslova N, Michna V, Kasny M, Mikel P, Kralik P. xMAP technology: applications in detection of pathogens. Front Microbiol. 2017;8:55.Google Scholar
  17. StephenAngeloni RC, Dunba S, Garcia C, Gibson G, Martin C, Stone V. xMAP Cookbook: A Collection of Methods and Protocols for Developing Multiplex Assays with xMAP Technology. 2nd ed; 2014.Google Scholar
  18. Opalka D, Lachman CE, MacMullen SA, Jansen KU, Smith JF, Chirmule N, Esser MT. Simultaneous quantitation of antibodies to neutralizing epitopes on virus-like particles for human papillomavirus types 6, 11, 16, and 18 by a multiplexed luminex assay 2. Clin Diagn Lab Immunol. 2003;10:108–15.PubMedPubMed CentralGoogle Scholar
  19. Dias D, Van Doren J, Schlottmann S, Kelly S, Puchalski D, Ruiz W, Boerckel P, Kessler J, Antonello JM, Green T, et al. Optimization and validation of a multiplexed Luminex assay to quantify antibodies to neutralizing epitopes on human papillomaviruses 6, 11, 16, and 18. Clin Diagn Lab Immunol. 2005;12:959–69.PubMedPubMed CentralGoogle Scholar
  20. Fonseca BP, Marques CF, Nascimento LD, Mello MB, Silva LB, Rubim NM, Foti L, Silva ED, Ferreira AG, Krieger MA. Development of a multiplex bead-based assay for detection of hepatitis C virus. Clin Vaccine Immunol. 2011;18:802–6.View ArticleGoogle Scholar
  21. Meulemans G, Halen P. Enzyme-linked immunosorbent assay (ELISA) for detecting infectious laryngotracheitis viral antibodies in chicken serum. Avian Pathol. 1982;11:361–8.View ArticleGoogle Scholar
  22. Nagano H, Tsuchimoto M, Hohdatsu T, Yamagami T, Ide S, Eiguchi Y, Tanaka Y, Yamagishi H. Enzyme-linked-immunosorbent-assay for the detection of infectious-bronchitis virus (Ibv) antigen with monoclonal-antibody. Jpn J Vet Sci. 1990;52:657–9.View ArticleGoogle Scholar
  23. Perrotta C, Furtek C, Wilson RA, Cowen BS, Eckroade RJ. A standardized enzyme-linked immunosorbent-assay for infectious-bronchitis virus - comparison with Hemagglutination-inhibition and virus-neutralization assays for measuring protective antibody-levels in chickens. Avian Dis. 1988;32:451–60.View ArticleGoogle Scholar
  24. Bokken GCAM, Bergwerff AA, van Knapen F. A novel bead-based assay to detect specific antibody responses against toxoplasma gondii and Trichinella spiralis simultaneously in sera of experimentally infected swine. BMC Vet Res. 2012;8:36.Google Scholar
  25. Baker HN, Murphy R, Lopez E, Garcia C. Conversion of a capture ELISA to a Luminex xMAP assay using a multiplex antibody screening method. J Vis Exp. 2012:4084.Google Scholar
  26. duPont NC, Wang KH, Wadhwa PD, Culhane JF, Nelson EL. Validation and comparison of luminex multiplex cytokine analysis kits with ELISA: determinations of a panel of nine cytokines in clinical sample culture supernatants. J Reprod Immunol. 2005;66:175–91.View ArticleGoogle Scholar
  27. Kong WW, Li Y, Cheng SH, Yan C, An SP, Dong Z, Yan LN, Yuan YH. Luminex xMAP combined with western blot improves HIV diagnostic sensitivity. J Virol Methods. 2016;227:1–5.View ArticleGoogle Scholar
  28. Zhang X, Zhao Y, Zhang X, Liu S. Preparation of polyclonal antibodies against gD protein of ILTV and application. J Northeast Agric Univ. 2016;5:8.Google Scholar
  29. He H, Xu M, Yu J, Wei X, Yan Y, Gao X, Liu X. Cloning and prokaryotic expression of nucleoprotein gene of infectious bronchitis virus (IBV) ZZ2004 strain. Chin Anim Husb Vet Med. 2014;41:15–9.Google Scholar
  30. Karanikola SN, Krucken J, Ramuenke S, de Waal T, Hoglund J, Charlier J, Weber C, Muller E, Kowalczyk SJ, Kaba J, et al. Development of a multiplex fluorescence immunological assay for the simultaneous detection of antibodies against Cooperia oncophora, Dictyocaulus viviparus and Fasciola hepatica in cattle. Parasit Vectors. 2015;8:335.Google Scholar

Copyright

© The Author(s). 2018

Advertisement