COMPARATIVE EVALUATION OF CULTURAL AND MOLECULAR IDENTIFICATION METHODS OF Mycoplasma pneumoniae FROM SPUTUM SAMPLES OF SUBJECTS IN NNEWI.

SOURCE:

Faculty: Health Sciences And Technology
Department: Medical Laboratory Science

CONTRIBUTORS:

Adike C. Ndidi
Agbakoba N.R.
Enweani I.B.

ABSTRACT:

Mycoplasma pneumoniae is an organism that belongs to the family mycoplasmataceae. It’s role as a disease causing agent continues to draw interest especially with the advent of highly sensitive detection techniques. This bacterium poses a health problem to both animals and humans resulting in serious illnesses such as community-acquired pneumonia, lung damage and death. The virulence gene of M. pneumoniae, glycerophosphodiesterase (GLPQ gene) has not been detected in Nigeria. This work investigated the prevalence of M. pneumoniae as agent of respiratory tract infections using culture and molecular methods of identification, in patients attending Pulmonary Tuberculosis Clinic at Nnamdi Azikiwe Teaching, Hospital, Nnewi as well as detecting the most virulence gene of this organism. A total of 263 sputum samples were collected: 188 test subjects and 75 control subjects. These samples were examined bacteriologically using PPLO broth and agar, MacConkey, blood and chocolate agars. M. pneumoniae was further identified by polymerase chain reaction (PCR) technique using M. pneumoniae specific primer. All M. pneumoniae DNA positive samples were further amplified to detect the GLPQ gene of the organism using GLPQ specific primer. The overall prevalence rates of M. pneumoniae among the 263 subjects were 4.9% and 8.0% by culture and PCR respectively. The prevalence rate of the organism was significantly higher among the test subjects 11(5.9%) by culture and 18(9.6%) by PCR than the control subjects 2(2.7%) by culture and 3(4.0%) by PCR. The colonization of the organism was significant among the age groups 31-40 years (P<0.05). The prevalence of M. pneumoniae most virulence gene (GLPQ) among the 18-PCR positive subjects was 27.8%. The GLPQ gene was detected only in the female gender and this showed that there was a significant difference between the occurrence of the GLPQ gene among the female gender than the male gender (P<0.05). The antibiotic sensitivity pattern of M. pneumoniae showed that the organism was susceptible to Lyntriaxone, Levofloxacin, Ciprofloxacin, Azithromycin and Doxycycline while it showed resistance to Septrin, Peflacine, Rifampicin, Erythromycin and Norbactin. Other bacteria isolated from the sputum specimens included Staphylococcus aureus, 35 (18.6%); Streptococcus pneumoniae, 31 (16.5%); Pseudomonas aeruginosa, 11(5.9%) and Klebsiella pneumoniae 12(6.4%). The presence of M. pneumoniae in the sputum samples of subjects as well as the presence of the virulence gene (GLPQ) only from test subjects shows that the bacterium actually contributed to the aetiology of lower respiratory tract infections. These were better detected by PCR. It is recommended that the search for M. pneumoniae be included in the routine laboratory investigations for lower respiratory tract infection

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