The application research of xTAG GPP multiplex PCR in the diagnosis of persistent and chronic diarrhea in children

Abstract Background Persistent and chronic diarrhea is difficult to treat, and infection is still the main causes. Clearly infected pathogens are essential for treatment. In this study, we investigate the application value of xTAG gastrointestinal pathogen panel (xTAG GPP) multiplex PCR in the early diagnosis of children with persistent and chronic diarrhea and understand the epidemiology of intestinal diarrhea pathogens. Methods One hundred and ninety-nine specimens were collected from Nanjing Children's Hospital Affiliated to Nanjing Medical University (Nanjing, China). Comparing the xTAG GPP multiplex PCR Assay with the traditional methods (culture, rapid enzyme immunoassay chromatography, microscopic examination) and madding the statistical analysis. Results The positive rate of 199 patients with diarrhea specimens was 72.86% (145/199). The virus detection rate was 48.7%, Rotavirus A was the most common organism detected (34.6%), concentrated in winter, popular in children. Secondly, Norovirus GI/GII (20.6%). The positive rate of bacteria was 40.2%, Campylobacter (22.11%, 44/199) was most frequently detected. With C. difficile toxins A/B and Salmonella detected 44 and 17 samples, respectively. Infections with Shigella occurred 4 times, E. coli O157 was only detected once. There were three samples with parasitic (1.51%), two samples were positive for Entamoeba histolytica, one for Cryptosporidium. Adenovirus40/41, STEC, ETEC, Giardia, Yersinia enterocolitica and Vibriocholera were not detected. Totally 86 (43.2%) infected specimens with single pathogen were detected. There were 57 co-infections (28.64% of samples) of viruses and/or bacteria and/or parasites. Co-infections involved 29 double infections (23.62%) samples, 9 triple infections (4.52%) and 2 quadruple infections (0.5%). Norovirus GI/GII was found to have the highest involvement in co-infections 30(15.08%). Conclusion xTAG GPP multiplex PCR is simple, sensitive, specific and can be used as a quick way to diagnose the children persistent and chronic diarrhea.

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A multiplex PCR-SSCP test to genotype bovine β-casein alleles A1 , A2 , A3 , B, and C

Animal Genetics ◽

10.1046/j.1365-2052.1999.00445-6.x ◽

1999 ◽

Vol 30 (4) ◽

pp. 322-323 ◽

Cited By ~ 9

Author(s):

A. Barroso ◽

S. Dunner ◽

J. Ca·ón

Keyword(s):

Multiplex Pcr

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Multiplex PCR zum Nachweis disseminierter Tumorzellen (DTC) im Blut von Patientinnen mit Mammakarzinom

Zentralblatt für Gynäkologie ◽

10.1055/s-2005-870717 ◽

2005 ◽

Vol 127 (03) ◽

Author(s):

J Rom ◽

A Schneeeweiss ◽

V Zieglschmid ◽

C Hollmann ◽

O Böcher ◽

...

Keyword(s):

Multiplex Pcr

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Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

Bangladesh Journal of Medical Microbiology ◽

10.3329/bjmm.v1i2.21506 ◽

2016 ◽

Vol 1 (2) ◽

pp. 38-42 ◽

Cited By ~ 4

Author(s):

Khairun Nessa ◽

Dilruba Ahmed ◽

Johirul Islam ◽

FM Lutful Kabir ◽

M Anowar Hossain

Keyword(s):

Escherichia Coli ◽

Multiplex Pcr ◽

Clinical Laboratory ◽

Proteinase K ◽

Microbiology Laboratory ◽

Pcr Assay ◽

E Coli ◽

Diarrheagenic Escherichia Coli ◽

Multiplex Pcr Assay ◽

Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

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