New York City medium for enhanced recovery of Mycoplasma pneumoniae from clinical specimens

Abstract

Modified New York City (MNYC) medium and PPLO medium without methylene blue (PPLO agar) were compared for their ability to support the growth of Mycoplasma pneumoniae from clinical specimens. Pharyngeal specimens were collected from 1,070 college students who visited the Syracuse University Student Health Center. Of these patients, 623 were symptomatic for respiratory infection, and the remaining 447 were asymptomatic for respiratory illness. Throat swabs were inoculated into PPLO broths, and these broths were subcultured onto MNYC medium and PPLO agar after 3 and 14 days of incubation. A total of 222 (20.7%) clinical isolates of M. pneumoniae were recovered on these solid media, with the majority of the isolates (196) recovered from symptomatic patients. All isolates grew on MNYC medium, whereas five isolates failed to grow on PPLO agar. All isolates of M. pneumoniae recovered from symptomatic patients were detected on MNYC medium within 1 to 5 days of incubation, whereas 5 to 7 days of incubation were required before mycoplasmal growth was detected on PPLO agar. Over 86% of these mycoplasma isolates were detected on MNYC medium within 3 days of incubation and before the detection of any mycoplasmal growth on PPLO agar. A similar pattern of recovery times was observed for mycoplasmas isolated from asymptomatic patients. The results of this study have shown that MNYC medium is better than PPLO agar in supporting the rapid growth of M. pneumoniae from clinical specimens after 72-h blind subculture in PPLO glucose broth.

The multivariate detection limit for Mycoplasma pneumoniae as determined by nanorod array-surface enhanced Raman spectroscopy and comparison with limit of detection by qPCR

“…Wild type M. pneumoniae strain M129 was used in this study. Mycoplasma samples were cultured in SP4 medium 3,22 in tissue culture asks with a 1 ml ml À1 inoculation, incubated at 37 C, and harvested at log phase when the phenol red indicator turned an orange color upon reaching a pH of $6.5. At time of harvest, spent growth medium was decanted and cells were scraped into 0.1Â volume of SP4.…”

Specificity and Strain-Typing Capabilities of Nanorod Array-Surface Enhanced Raman Spectroscopy for Mycoplasma pneumoniae Detection

“…P1 genotype groups were determined by the Pneumonia Response and Surveillance Laboratory at the CDC in Atlanta, GA using DNA sequence analysis, qPCR in combination with high resolution melt curve analysis, and RFLP sequencing analysis. All mycoplasma isolates and controls were cultured in SP4 medium [ 2 , 30 ] in tissue culture flasks with a 1μl/ml inoculation and incubated at 37°C. Strains grown at UGA were harvested at log phase when the phenol red indicator turned an orange color upon reaching a pH of ~6.5.…”

“…The highly reproducible detection capabilities of NA-SERS have been well demonstrated for multiple infectious agents, including RSV, rotavirus, influenza, HIV, adenovirus, SARS coronavirus, and M . pneumoniae [ 13 , 22 , 28 – 30 ].…”

Comparison of the Efficacy of Egg Yolk Extract and Horse Serum for Growth of Mycoplasma pneumoniae

“…As the growth of Mycoplasma pneumoniae is often markedly affected by certain HS preparations, it is important to use HS preparations of suitable quality in the medium for diagnosis of M. pneumoniae infection. Recently SP-4 medium, which contains fetal bovine serum heated at 56 C for 1 hr (11), and modified New York City medium, which contains agamma HS (4,5), have been introduced as excellent primary isolation media for M. pneumoniae.…”