Development of a novel DNA microarray to detect bacterial pathogens in patients with chronic obstructive pulmonary disease (COPD).
A novel microarray was constructed with DNA PCR product probes targeting species specific functional genes of nine clinically significant respiratory pathogens, including the Gram-positive organisms (Streptococcus pneumoniae, Streptococcus pyogenes), the Gram-negative organisms (Chlamydia pneumoniae, Coxiella burnetii Haemophilus spp., Legionella pneumophila, Moraxella catarrhalis, and Pseudomonas aeruginosa), as well as the atypical bacterium, Mycoplasma pneumoniae. In a "proof-of-concept" evaluation of the developed microarray, the microarray was compared with real-time PCR from 14 sputum specimens from COPD patients. All of the samples positive for bacterial species in real-time PCR were also positive for the same bacterial species using the microarray. This study shows that a microarray using PCR probes is a potentially useful method to monitor the populations of bacteria in respiratory specimens and can be tailored to specific clinical needs such as respiratory infections of particular patient populations, including patients with cystic fibrosis and bronchiectasis.
Curran
Tanya
T
Microbiology Department, Kelvin Building, The Royal Group of Hospitals and Dental Hospital Health and Social Services Trust, Belfast, BT12 6BA, UK. tanya.curran@belfasttrust.hscni.net
Coulter
Wilson A
WA
Fairley
Derek J
DJ
McManus
Terence
T
Kidney
Joe
J
Larkin
Mike
M
Moore
John E
JE
Coyle
Peter V
PV
D016993:Chlamydophila pneumoniae / Q000235:genetics / Q000302:isolation & purification; D016997:Coxiella burnetii / Q000235:genetics / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D006090:Gram-Negative Bacteria / Q000235:genetics / Q000302:isolation & purification*; D016905:Gram-Negative Bacterial Infections / Q000175:diagnosis* / Q000209:etiology; D016908:Gram-Positive Bacterial Infections / Q000175:diagnosis* / Q000209:etiology; D006190:Haemophilus / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D016952:Legionella pneumophila / Q000235:genetics / Q000302:isolation & purification; D001936:Moraxella catarrhalis / Q000235:genetics / Q000302:isolation & purification; D009177:Mycoplasma pneumoniae / Q000235:genetics / Q000302:isolation & purification; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000302:isolation & purification; D029424:Pulmonary Disease, Chronic Obstructive / Q000150:complications / Q000382:microbiology*; D012141:Respiratory Tract Infections / Q000175:diagnosis* / Q000209:etiology; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013183:Sputum / Q000382:microbiology; D013296:Streptococcus pneumoniae / Q000235:genetics / Q000302:isolation & purification*; D013297:Streptococcus pyogenes / Q000235:genetics / Q000302:isolation & purification*
D004269:DNA, Bacterial