关键词: 钩端螺旋体, 聚合酶链式反应, 16S rRNA

Abstract: Objective To compare and evaluate the detection effect between an improved polymerase chain reaction (PCR) method and three current standard methods for pathogenic Leptospira, and to provide an improved method in early diagnosis of patients and host animal monitoring. Methods A step of reverse transcription process was added to the recommended pathogenic Leptospira detection method (method 3) in National Vector Etiology Surveillance Program (Trial), without changing the primers and probes, to form an improved method (method 4). Four PCR methods, i.e., Method 4, the recommended method of Industry Standards for Health Care (method 1), the recommended method of Industry Standards for Inspection and Quarantine (method 2), and method 3, were used to detect Leptospira positive samples with series dilutions, early serum samples of patients, and rodent kidney specimens in parallel, and to evaluate the detection effect. Results The sensitivity of method 4 was 10 000 times that of methods 1 and 2, and 1 000 times that of method 3. Leptospira nucleic acids in early serum of patients were only detected by method 4. The detection rates of Leptospira in 100 rodent kidney specimens were 2.00%, 4.00%, 8.00%, and 40.00% by the four methods, respectively. There was a significant difference in the positive detection rate between method 4 and methods 1, 2, and 3 (all P<0.001) and no significant difference between methods 1, 2, and 3 (P>0.05). Conclusion The improved method has a superior detection effect to the other three current standard methods in the early diagnosis of patients with leptospirosis and the rodent kidney.

Key words: Leptospira, Polymerase chain reaction, 16S rRNA