Objective: To study the effects of antibiotic treatment time limit for children with purulent meningeal cerebrospinal fluid bacterial 16s rRNA genes detection. Methods: From February 2014 to March 2016, selected 60 cases of patients with suppurative meningeal cerebrospinal fluid in our hospital. All children were given 16s rRNA gene detection and cerebrospinal fluid bacterial culture, compared the results of the two methods, and observed the effects of antibiotic on two kinds of detection methods. Results: The positive rate of CSF germiculture was significantly lower than that of 16s rRNA gene chip testing [15% (9/60) vs. 35% (21/60)] (P<0.05). 16s rRNA genes detection positive rate of early and late antibiotic application had no significant difference [36.67% (11/30) vs. 33.33% (10/30)] (P>0.05); cerebrospinal fluid germiculture positive rate in early antibiotic application were higher than that in late antibiotic application [13.33% (4/30) vs. 0.00% (0/30)] (P<0.05). Conclusion: Compared with CSF cultures, 16s rRNA gene chip method detection in cerebrospinal fluid bacteria of suppurative meningeal children is less affected by pmin of antibiotic treatment.