Construction and Application of MALDI-TOF Mass Spectrometry for the Detection of Haemophilus parasuis.

Xiaoxu Wang,Feng Xu,Kun Ning,Xinyong Qi,Liping Shen,Jian Wang,Junyan Liu

doi:10.1155/2021/5588855

Abstract

To construct a protein fingerprint database of Haemophilus parasuis (H. parasuis), thus improving its clinical diagnosis efficiency. A total of 15 H. parasuis standard strains were collected to establish a protein fingerprint database of H. parasuis using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), and the effects of different culture media and culture time on the quality and identification results of the protein fingerprint were investigated. The results showed that tryptone soy agar (TSA) and tryptone soy broth (TSB) media and different incubation times had no significant effect on the characteristic peaks of the protein profiles. In addition, 18 clinical isolates were used to compare the identification results of the self-built protein fingerprint database, PCR detection, and basic database. Only one strain was identified in the original VITEK-MS system database, while the self-made protein fingerprint database of H. parasuis was 100% accurate for the detection of 18 clinical isolate strains. The protein fingerprint database of H. parasuis built by our laboratory is suitable for rapid clinical diagnosis of H. parasuis, due to its high accuracy, efficiency, and strong specificity.

Highlights

With the improvement of the Chinese economy, the pig industry has reached a great development, so how to produce commercial pigs in a healthy and efficient way has become the key
The mass spectra of H. parasuis standard strain CVCC3892 inoculated in tryptone soy agar (TSA) and tryptone soy broth (TSB) media were compared
The result showed no significant difference in the characteristic peaks between the two (Figure 1), indicating that the culture medium had no significant effect on the characteristic peaks of the protein mass spectra

Summary

Introduction

With the improvement of the Chinese economy, the pig industry has reached a great development, so how to produce commercial pigs in a healthy and efficient way has become the key. Haemophilus parasuis (H. parasuis) disease, known as Glässer’s disease [1], is induced by H. parasuis, leading to the increase of body temperature, joint swelling, dyspnea, polyserositis, and arthritis in pigs [2]. It is an infectious disease with a high mortality rate, seriously endangering the health of piglets and young pigs. H. parasuis is a Gram-negative, nonmotile, small, pleomorphic bacterium belonging to genus Haemophilus in the family Pasteurellaceae [3]. Virulent strains can as secondary microorganism of pneumonia cause septicemia without polyserositis or Glässer’s disease characterized by polyserositis, pericarditis, arthritis, and meningitis [5]

Methods

Results

Conclusion