Kumar Yashwant, Mani Kavaratty Raju, Tahlan Ajay Kumar
National Salmonella and Escherichia Centre, Central Research Institute, Kasauli-173204, Himachal Pradesh, India.
Trop Life Sci Res. 2019 Jan;30(1):57-71. doi: 10.21315/tlsr2019.30.1.4. Epub 2019 Jan 31.
A number of countries, including developed countries, still have typhoid fever as a major problem resulting in frequent outbreaks. The importance of controlling spread of typhoid fever is well known and necessitates periodic studies to delineate epidemiological relationships. Although phage typing remains to be the preferred conventional method for characterisation of typhoid bacilli, it is of limited use due to prevalence of few predominant phage types in the country like India. Therefore, an effort has been made to assess three molecular methods [Outer Membrane Protein (OMP) Profiling, Random Amplification of Polymorphic DNA (RAPD) and Pulsed Field Gel Electrophoresis (PFGE)] for typing of serovar Typhi. 128 serovar Typhi isolates were identified using biotyping and serotyping followed by antimicrobial susceptibility testing. These isolates were further subjected to OMP analysis, RAPD and PFGE. PFGE (114 unique clusters) was found to be the most discriminatory method followed by RAPD (94 unique clusters) and OMP profiling (50 unique clusters). Multidrug resistant strains were well discriminated by all three methods used in the study. PFGE still remains the most preferred method for detailed epidemiological investigations. However, random amplification of polymorphic DNA and outer membrane protein profiling can also be considered for molecular discrimination of the isolates in the laboratories lacking high-end facilities.
包括发达国家在内的许多国家,伤寒热仍是一个导致频繁暴发的主要问题。控制伤寒热传播的重要性众所周知,因此需要定期开展研究以明确流行病学关系。尽管噬菌体分型仍然是鉴定伤寒杆菌的首选传统方法,但由于在印度等国家少数几种主要噬菌体类型较为普遍,其用途有限。因此,已尝试评估三种分子方法[外膜蛋白(OMP)分析、随机扩增多态性DNA(RAPD)和脉冲场凝胶电泳(PFGE)]用于伤寒血清型的分型。通过生物分型和血清分型鉴定出128株伤寒血清型菌株,随后进行药敏试验。这些菌株进一步进行OMP分析、RAPD和PFGE检测。发现PFGE(114个独特聚类)是最具鉴别力的方法,其次是RAPD(94个独特聚类)和OMP分析(50个独特聚类)。研究中使用的所有三种方法都能很好地区分多重耐药菌株。PFGE仍然是详细流行病学调查的最首选方法。然而,在缺乏高端设备的实验室中,随机扩增多态性DNA和外膜蛋白分析也可用于菌株的分子鉴别。