Identification of Candida Spp. Isolated From Urine by Phenotypic Methods and Multiplex Pcr in Duhok, Iraq

Introduction: Community and healthcare associated infections caused by multi-drug resistant gram negative organisms (MDR GN) represent a worldwide threat. Nucleic Acid Detection tests are becoming more common for their detection; however they can be expensive requiring specialised equipment and local expertise. This study was done to evaluate the utility of a commercial multiplex tandem (MT) PCR for detection of MDR GN. Methods: The study was done on stored laboratory MDR GN isolates from sterile and non-sterile specimens (n=126, out of stored 567 organisms). Laboratory validation of the MT PCR was done to evaluate sensitivity, specificity and agreement with the current phenotypic methods used in the laboratory. Amplicon sequencing was also done on selected isolates for assessing performance characteristics. Workflow and cost implications of the MT PCR were evaluated. Results: The sensitivity and specificity of the MT PCR were calculated to be 95% and 96.7% respectively. Agreement with the phenotypic methods was 80%. Major lack of agreement was seen in detection of AmpC beta lactamase in enterobacteriaceae and carbapenemase in non-fermenters. Agreement of the MT PCR with another multiplex PCR was found to be 87%. Amplicon sequencing confirmed the genotype detected by MT PCR in 94.2 % of cases tested. Time to result was faster for the MT PCR but cost per test was higher. Conclusion: This study shows that with carefully chosen targets for detection of resistance genes in MDR GN, rapid and efficient identification is possible. MT PCR was sensitive and specific and likely more accurate than phenotypic methods.

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Detection of carbapenem resistant enterobacteriaecae and the comparison between phenotypic methods and multiplex PCR

Indian Journal of Microbiology Research ◽

10.18231/j.ijmr.2019.027 ◽

2019 ◽

Vol 6 (2) ◽

pp. 126-130

Author(s):

Rubhana Qadri ◽

◽

Suhail Ahmed ◽

Dalip K Kakru ◽

Gulnaz Bashir ◽

...

Keyword(s):

Multiplex Pcr ◽

Carbapenem Resistant ◽

Phenotypic Methods

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The use of multiplex-PCR method in identification of Candida species from vaginal candidiasis patients

Biodiversitas Journal of Biological Diversity ◽

10.13057/biodiv/d201040 ◽

2019 ◽

Vol 20 (10) ◽

Author(s):

SUSILAWATI ◽

KEMAS YA’KUB RAHADIYANTO ◽

MUHAIMIN RAMDJA ◽

THEODORUS ◽

Hermansyah Hermansyah

Keyword(s):

Multiplex Pcr ◽

Candida Species ◽

High Sensitivity ◽

High Specificity ◽

Vaginal Swab ◽

Vaginal Candidiasis ◽

Candida Spp ◽

Universal Primer ◽

Drug Of Choice ◽

Pcr Method

Abstract. Susilawati, Rahadiyanto KY, Ramdja, Theodorus, Hermansyah. 2019. The use of multiplex-PCR method in identification of Candida species from vaginal candidiasis patients. Biodiversitas 20: 3063-3069. Vaginal Candidiasis is an infection caused by a yeast called Candida which may be resulted in different virulences, thus leading to a varied drug of choice for therapy. The objective of this study was to determine the sensitivity and specificity of multiplex-PCR test in detecting Candida species in women with clinical diagnosis of vaginal candidiasis. There were 79 vaginal swab samples which fulfill inclusion criteria. Samples were cultured at ASD media for isolation of Candida spp. Identification of Candida spp. was conducted using both biochemical fermentation and multiplex-PCR methods. Primers pairs used in multiplex-PCR were universal primer ITS1 and ITS2, and specific primer CA3 and CA4. The identification test of multiplex-PCR resulted in Candida krusei: sensitivity: 100%, specificity:61,1%, Positive Prediction Value (PPV):63.2%, Negative Prediction Value (NPV): 100%; C. glabrata: sensitivity:0%, specificity: 100% PPV: 0% NPV: 76.7%; C. tropicalis: sensitivity:100%, spesificity:100%, PPV: 100% NPV: 100%; C. albicans: sensitivity: 33.3%, specificity: 100%, PPV: 100%, NPV: 93.1% and C. stellatoidea: sensitivity:: 100%, specificity:92.6%, PPV: 60%, NPV: 100%. These results suggested that multiplex-PCR method becomes a rapid alternative in identifying species of Candida due to its high sensitivity and high specificity.

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