Context
Pneumonia is an illness of the lungs and respiratory system in which the alveoli become inflamed and flooded with fluid. It is a common illness, occurs in all age groups and is a leading cause of death in immunocompromised patients. Pneumonia can result from a variety of causes, including infection with bacteria, viruses, fungi, or parasites. Some bacteria that cause pneumonia include Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa and Moraxella catarrhalis. "Atypical" bacteria which cause pneumonia include Chlamydophila pneumoniae, Mycoplasma pneumoniae, and Legionella pneumophila.
Typical symptoms associated with pneumonia include cough, chest pain, fever, and difficulty breathing. Treatment depends on the cause of pneumonia, bacterial pneumonia is treated with antibiotics. Because treatment should generally not be delayed in any person with a serious pneumonia, empiric treatment is usually started well before laboratory reports are available.
Diagnostic tools include x-rays, physical examination, examination of the sputum and blood tests. It is not possible to clearly determine the microbiologic cause of a pneumonia based on x-rays alone. And laboratory testing typically takes several days so microbiologic classification is usually not possible at the time of initial diagnosis. Diagnosing pneumonia can be difficult in some people, especially those who have other illnesses.
Approach
Pneumonia might be diagnosed by the characterisation of markers proteins in patient sera.
Functional domains of the target proteins were annotated by specialists from the ISS. The use of appropriate Bioinformatic tools allowed to constitute the starting list of proteins targets.
At the beginning of this work, 80 proteins from L. pneumophila and 98 proteins from C. pneumoniae were selected. On the whole, 183 genes went through a parallel cloning, expression screening and micro-purification protocol.
70 proteins were produced and purified (41 from C. pneumoniae, 28 from L. pneumophila).
33 target proteins, 18 from C. pneumoniae and 15 from L. pneumophila were validated after an immune response in presence of patient sera.
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