Multiplex PCR for Diagnosis of Salmonella enterica Serovar Typhi

被引:9
作者
El-Sayed, Ahmed K. A. [1 ]
El-Shishtawy, Mamdouh [2 ]
El-Taweel, Fathy [3 ]
El-Mansoury, Heba [3 ]
机构
[1] Damietta Univ, Fac Sci, Dept Bot, Damietta El Gededa, Egypt
[2] Mansoura Univ, Fac Pharm, Dept Biochem, Mansoura, Egypt
[3] Damietta Univ, Fac Sci, Dept Chem, Damietta El Gededa, Egypt
关键词
Salmonella Typhi; Widal test; culturing; invA; fliC; POLYMERASE-CHAIN-REACTION; NESTED PCR; WIDAL TEST; FEVER; IDENTIFICATION; SAMPLES; BLOOD; AGAR; MEDIA;
D O I
10.7754/Clin.Lab.2015.150115
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
Background: Typhoid fever caused by Salmonella enterica serovar Typhi is endemic in developing countries. Its detection by culturing on specific media or serological methods like the Widal test is time consuming and/or less sensitive than the PCR technique. Here we apply a multiplex PCR dependent detection for S. Typhi using two specific primers for invA and fliC genes. Methods: 76 patients with clinical suspicion of typhoid fever were examined. Two sets of primers derived from invA and fliC genes specific for Salmonella spp. and S. Typhi were used as multiplex PCR in order to detect the pathogen in their blood samples. This was compared with traditional culturing methods on two different chromogenic media [Melibiose, mannitol, and sorbitol (MMS) agar media specific for S. Typhi and Salmonella-Shigella (SS) agar media]. Also, the suspected typhoid samples were tested by Widal 0 antigen. Results: The sensitivities of culturing, Widal test, and multiplex PCR were 61.36%, 88.64%, and 100%, respectively, while their specificities were 100%, 62.50%, and 86.49%, respectively. The multiplex PCR showed higher efficiency reaching 93.42% compared to culturing and Widal test which was 77.63%. Conclusions: The high sensitivity, specificity, and efficiency of the studied multiplex PCR encourage us to recommend it as a useful tool for S. Typhi detection not only in clinically suspected negative culture individuals, but also for the false positive Widal test cases of typhoid fever in endemic regions.
引用
收藏
页码:1537 / 1543
页数:7
相关论文
共 40 条
[1]   Diagnosis of typhoid fever by polymerase chain reaction [J].
Ambati, S. R. ;
Nath, Gopal ;
Das, B. K. .
INDIAN JOURNAL OF PEDIATRICS, 2007, 74 (10) :909-913
[2]  
Ausubel FM, 2003, CURRENT PROTOCOL S59
[3]   Development of a Multiplex Primer Extension Assay for Rapid Detection of Salmonella Isolates of Diverse Serotypes [J].
Ben-Darif, Elloulu ;
Jury, Francine ;
De Pinna, Elizabeth ;
Threlfall, E. John ;
Bolton, Frederick J. ;
Fox, Andrew J. ;
Upton, Mathew .
JOURNAL OF CLINICAL MICROBIOLOGY, 2010, 48 (04) :1055-1060
[4]   Typhoid and paratyphoid fever [J].
Bhan, MK ;
Bahl, R ;
Bhatnagar, S .
LANCET, 2005, 366 (9487) :749-762
[5]   Current concepts in the diagnosis and treatment of typhoid fever [J].
Bhutta, Zulfiqar A. .
BMJ-BRITISH MEDICAL JOURNAL, 2006, 333 (7558) :78-82B
[6]   Standardisation of polymerase chain reaction for the detection of Salmonella typhi in typhoid fever [J].
Chaudhry, R ;
Laxmi, BV ;
Nisar, N ;
Ray, K ;
Kumar, D .
JOURNAL OF CLINICAL PATHOLOGY, 1997, 50 (05) :437-439
[7]   Rapid identification of Salmonella serovars in feces by specific detection of virulence genes, invA and spvC, by an enrichment broth culture-multiplex PCR combination assay [J].
Chiu, CH ;
Ou, JT .
JOURNAL OF CLINICAL MICROBIOLOGY, 1996, 34 (10) :2619-2622
[8]   COMPARISON OF RAMBACH AGAR, SM-ID MEDIUM, AND HEKTOEN ENTERIC AGAR FOR PRIMARY ISOLATION OF NONTYPHI SALMONELLAE FROM STOOL SAMPLES [J].
DUSCH, H ;
ALTWEGG, M .
JOURNAL OF CLINICAL MICROBIOLOGY, 1993, 31 (02) :410-412
[9]   LYSINE-IRON AGAR IN DETECTION OF ARIZONA CULTURES [J].
EDWARDS, PR ;
FIFE, MA .
APPLIED MICROBIOLOGY, 1961, 9 (06) :478-&
[10]   DETECTION OF SALMONELLA-TYPHI BY PCR [J].
FRANKEL, G .
JOURNAL OF CLINICAL MICROBIOLOGY, 1994, 32 (05) :1415-1415