Wolf J P, Marcus L
Appl Microbiol. 1969 Jul;18(1):4-7. doi: 10.1128/am.18.1.4-7.1969.
An impediment to progress in the study of the course of growth, the effects of medium components, antibiotics, etc., of Mycoplasma has been the cumbersome methods of growth measurement currently in use. Heretofore, it required the plating of numerous samples during growth, at least in triplicate, after appropriate dilution, followed by a delay of 2 to 3 days before the colonies developed so that counts could be made. We applied the technique of light scattering to measure the growth of Mycoplasma laidlawii in liquid culture continuously in a manner analogous to the use of absorbancy for bacteria. Scattered light measurements precisely paralleled data obtained by the tedious method of plate counts and were available immediately during the development of the culture. The lower limit of sensitivity with the system described is 10(5)Mycoplasma per ml. The presence of serum in the medium lowers sensitivity somewhat. However, concentrations of serum up to 10% are easily tolerated. Higher serum content may require calibration curves. Thus the technique may be used with many pathogens, etc., that require serum to develop. One can easily and rapidly measure differences in growth rates as well as final cell yields during the course of growth, rather than 3 days later, after colonies have developed.
支原体生长过程、培养基成分、抗生素等影响因素研究进展的一个阻碍是目前使用的繁琐的生长测量方法。在此之前,在生长过程中需要对大量样品进行接种,至少一式三份,经过适当稀释后,在菌落形成前要延迟2至3天才能进行计数。我们应用光散射技术,以类似于使用吸光度测量细菌生长的方式,连续测量液体培养物中莱氏无胆甾原体的生长。散射光测量结果与通过繁琐的平板计数法获得的数据精确平行,并且在培养过程中可立即获得。所述系统的灵敏度下限为每毫升10(5)个支原体。培养基中血清的存在会使灵敏度有所降低。然而,高达10%的血清浓度很容易耐受。血清含量更高时可能需要校准曲线。因此,该技术可用于许多需要血清才能生长的病原体等。人们可以轻松快速地测量生长过程中的生长速率差异以及最终细胞产量,而不是在菌落形成3天后。