درمان آلودگي مايكوپلاسمايي در كشت سلول با سيپرو فلوكساسين و انروفلوكساسين: گزارش كوتاه

Mycoplasma contamination in cell cultures treated with ciprofloxacin and enrofloxacin: brief report

زمينه و هدف: در اين مطالعه رده‌های سلولی Vero آلوده به مايكوپلاسما، با رقت‌های مختلف سيپروفلوكساسين و انروفلوكساسين كه مانع همانندسازی DNA و سنتز پروتيين می‌شوند، تحت درمان قرار گرفتند. روش بررسی: در اين پژوهش اكتشافی كه در آزمايشگاه پژوهش و ساخت واكسن هاری انسانی انستيتو پاستور از آبان 1392 تا خرداد 1393انجام گرديد، رقت‌های مختلف آنتی‌بيوتيكی مورد استفاده و مقايسه قرار گرفت. عملكرد اين روش درمانی با PCR ارزيابی شد. با استفاده از روش تاپسيس (Technique for Order of Preference by Similarity to Ideal Solution, TOPSIS) كه يك روش تصميم‌گيری چند جانبه است بهترين اثر غلظت بر زمان به‌دست‌آمده در درمان مايكوپلاسما تعيين شد. يافته‌ها: بهترين غلظت به‌دست‌آمده جهت درمان مايكوپلاسمايی به‌ترتيب 20 و μg/ml 200 از سيپروفلوكساسين و برای درمان سلول‌های آلوده با انروفلوكساسين به‌ترتيب 3 و 30 و μg/ml 300 بود. نتيجه‌گيری: در مطالعه حاضر درمان خط سلولی Vero آلوده به مايكوپلاسما با آنتی‌بيوتيك‌های سيپروفلوكساسين و انروفلوكساسين بدون نياز به تغيير آنتی‌بيوتيك، كه در ساير فرايندها معمول است، انجام شد.

Background: Mycoplasma contamination in cell cultures is considered as a major economic, research and production problem. In this study, mycoplasma-infected Vero cell lines were treated by various dilutions of ciprofloxacin and enrofloxacin in a timely manner. Removal of mycoplasma contamination from infected cell cultures was evaluated and demonstrated by polymerase chain reaction (PCR) method. Methods: This study was done from October 2013 to May 2014, in Human Rabies Vaccine Laboratory, Pasteur Institute Production and Research Complex, Tehran, Iran. Different dilutions of ciprofloxacin and enrofloxacin were used in sequential passages for treatment of infected Vero cell line. Based on lowest passages of the cell line, antibiotic treatment with ciprofloxacin and enrofloxacin was done. Amelioration of the infection and removal of mycoplasma contamination was confirmed in each step by PCR method. The technique for order of preference by similarity to ideal solution, TOPSIS method, was used to suggest the most efficient concentration of ciprofloxacin and enrofloxacin. Results: Proposed concentration of ciprofloxacin is 20 μg/ml, and in the second order is 200 μg/ml. For enrofloxacin the best proposed concentrations are 30, 300 and 3 μg/ml respectively. Ciprofloxacin and enrofloxacin and ability of them for removal of mycoplasma and also the time of treatment were verified by evaluation of the recurrence of infection through consecutive subcultures of the treated cell line. Conclusion: Our results showed that 20 μg/ml of ciprofloxacin was the dilution of choice for mycoplasma elimination followed by 200 μg/ml of ciprofloxacin. Concentrations of 3, 30 and 300 of enrofloxacin, respectively, are appropriate for mycoplasma removal. More detailed works would be needed to verify the authenticity of the proposed simple and affordable way of mycoplasma elimination.