Comparative Diagnostic Techniques for Cryptosporidium Infection

Diarrhoea caused by Cryptosporidium is usually mild in immune competent individuals but severe in the young and those with underlying disease leading to compromised immunity. The conventional diagnosis of Cryptosporidium requires observation of the infective oocysts however, their tiny size yields i...

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Main Authors: Beauty E. Omoruyi, Uchechukwu U. Nwodo, Chukwuneke S. Udem, Francis O. Okonkwo
Format: Article
Language:English
Published: MDPI AG 2014-02-01
Series:Molecules
Subjects:
PCR
Online Access:http://www.mdpi.com/1420-3049/19/2/2674
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spelling doaj-c8262cc35c004342b166d0a3746b71272020-11-24T20:56:18ZengMDPI AGMolecules1420-30492014-02-011922674268310.3390/molecules19022674molecules19022674Comparative Diagnostic Techniques for Cryptosporidium InfectionBeauty E. Omoruyi0Uchechukwu U. Nwodo1Chukwuneke S. Udem2Francis O. Okonkwo3Department of Biochemistry and Microbiology, University of Forth Hare, Private Bag X1314, Alice 5700, South AfricaDepartment of Biochemistry and Microbiology, University of Forth Hare, Private Bag X1314, Alice 5700, South AfricaDepartment of Veterinary Physiology and Pharmacology, University of Nigeria, Nsukka 410002, NigeriaDepartment of Biochemistry, University of Nigeria, Nsukka 410002, NigeriaDiarrhoea caused by Cryptosporidium is usually mild in immune competent individuals but severe in the young and those with underlying disease leading to compromised immunity. The conventional diagnosis of Cryptosporidium requires observation of the infective oocysts however, their tiny size yields indistinct results, thus limiting the effectiveness of the conventional diagnostic technique, modified Ziehl-Neelsen (ZN) differential staining. Consequent to the abovementioned limitation, ZN staining, sandwich antigen detection enzyme linked immunosorbent assay (sad-ELISA) and a direct polymerase chain reaction (PCR) assay techniques were evaluated for diagnostic efficacy. Stool samples were collected from 180 consenting adult patients attending outpatient and inpatient clinics at Victoria Hospital, Alice, Eastern Cape Province of South Africa. Subjects were stratified as; 35 HIV-positive and diarrhoeagenic, 125 HIV-negative diarrhoeagenic and 20 apparently healthy controls. Cryptosporidium incidence following diagnostic techniques were 13 (37.1%; ZN staining), 26 (74.3%; sad-ELISA) and 23 (65.7%; PCR), respectively, among HIV-positive diarrhoeagenic patients and 34 (27.2%; ZN staining), 96 (76.8%; sad-ELISA) and 89 (71.2%; PCR) among HIV-negative diarrhoeagenic patients. Sensitivity, specificity and predictive values of the diagnostic techniques’ efficiency were: sensitivity: 46.2% (HIV-positive) and 32.3% (HIV-negative) against the ZN technique and 96.9% against sad-ELISA and PCR, respectively, for both HIV-positive and -negative patients; specificity was 88.9% (HIV-positive) and 96.6% (HIV-negative) against the ZN technique. Lastly, the predictive values were 92.3% (HIV-positive) and 96.9% (HIV-negative), respectively, following ZN staining. The sad-ELISA technique proved more suitable for the determination of the presence of Cryptosporidium oocysts. The high incidence of Cryptosporidium in HIV-positive subjects as compared to the HIV-negative population accentuates the significance of cryptosporidiosis diagnosis in the treatment and management of HIV cases.http://www.mdpi.com/1420-3049/19/2/2674cryptosporidiosisCryptosporidium speciesstaining techniquesantigen detection ELISAPCR
collection DOAJ
language English
format Article
sources DOAJ
author Beauty E. Omoruyi
Uchechukwu U. Nwodo
Chukwuneke S. Udem
Francis O. Okonkwo
spellingShingle Beauty E. Omoruyi
Uchechukwu U. Nwodo
Chukwuneke S. Udem
Francis O. Okonkwo
Comparative Diagnostic Techniques for Cryptosporidium Infection
Molecules
cryptosporidiosis
Cryptosporidium species
staining techniques
antigen detection ELISA
PCR
author_facet Beauty E. Omoruyi
Uchechukwu U. Nwodo
Chukwuneke S. Udem
Francis O. Okonkwo
author_sort Beauty E. Omoruyi
title Comparative Diagnostic Techniques for Cryptosporidium Infection
title_short Comparative Diagnostic Techniques for Cryptosporidium Infection
title_full Comparative Diagnostic Techniques for Cryptosporidium Infection
title_fullStr Comparative Diagnostic Techniques for Cryptosporidium Infection
title_full_unstemmed Comparative Diagnostic Techniques for Cryptosporidium Infection
title_sort comparative diagnostic techniques for cryptosporidium infection
publisher MDPI AG
series Molecules
issn 1420-3049
publishDate 2014-02-01
description Diarrhoea caused by Cryptosporidium is usually mild in immune competent individuals but severe in the young and those with underlying disease leading to compromised immunity. The conventional diagnosis of Cryptosporidium requires observation of the infective oocysts however, their tiny size yields indistinct results, thus limiting the effectiveness of the conventional diagnostic technique, modified Ziehl-Neelsen (ZN) differential staining. Consequent to the abovementioned limitation, ZN staining, sandwich antigen detection enzyme linked immunosorbent assay (sad-ELISA) and a direct polymerase chain reaction (PCR) assay techniques were evaluated for diagnostic efficacy. Stool samples were collected from 180 consenting adult patients attending outpatient and inpatient clinics at Victoria Hospital, Alice, Eastern Cape Province of South Africa. Subjects were stratified as; 35 HIV-positive and diarrhoeagenic, 125 HIV-negative diarrhoeagenic and 20 apparently healthy controls. Cryptosporidium incidence following diagnostic techniques were 13 (37.1%; ZN staining), 26 (74.3%; sad-ELISA) and 23 (65.7%; PCR), respectively, among HIV-positive diarrhoeagenic patients and 34 (27.2%; ZN staining), 96 (76.8%; sad-ELISA) and 89 (71.2%; PCR) among HIV-negative diarrhoeagenic patients. Sensitivity, specificity and predictive values of the diagnostic techniques’ efficiency were: sensitivity: 46.2% (HIV-positive) and 32.3% (HIV-negative) against the ZN technique and 96.9% against sad-ELISA and PCR, respectively, for both HIV-positive and -negative patients; specificity was 88.9% (HIV-positive) and 96.6% (HIV-negative) against the ZN technique. Lastly, the predictive values were 92.3% (HIV-positive) and 96.9% (HIV-negative), respectively, following ZN staining. The sad-ELISA technique proved more suitable for the determination of the presence of Cryptosporidium oocysts. The high incidence of Cryptosporidium in HIV-positive subjects as compared to the HIV-negative population accentuates the significance of cryptosporidiosis diagnosis in the treatment and management of HIV cases.
topic cryptosporidiosis
Cryptosporidium species
staining techniques
antigen detection ELISA
PCR
url http://www.mdpi.com/1420-3049/19/2/2674
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