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Original Article

Prevalence of Cryptosporidium-Associated Diarrhea in a High Altitude-Community of Saudi Arabia Detected by Conventional and Molecular Methods

Yousry Hawash1,2, Laila Sh. Dorgham3,4, Ayman S. Al-Hazmi2, Mohammed S. Al-Ghamdi2
The Korean Journal of Parasitology 2014;52(5):479-485.
Published online: October 22, 2014

1Department of Medical Parasitology, NLI, Menoufia University, Shebin El-Koom, Menoufia, Egypt.

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Corresponding author (yousryhawash@gmail.com)
• Received: April 28, 2014   • Revised: June 26, 2014   • Accepted: July 15, 2014

© 2014, Korean Society for Parasitology and Tropical Medicine

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Prevalence of Cryptosporidium-Associated Diarrhea in a High Altitude-Community of Saudi Arabia Detected by Conventional and Molecular Methods
Korean J Parasitol. 2014;52(5):479-485.   Published online October 22, 2014
DOI: https://doi.org/10.3347/kjp.2014.52.5.479
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Prevalence of Cryptosporidium-Associated Diarrhea in a High Altitude-Community of Saudi Arabia Detected by Conventional and Molecular Methods
Korean J Parasitol. 2014;52(5):479-485.   Published online October 22, 2014
DOI: https://doi.org/10.3347/kjp.2014.52.5.479
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Prevalence of Cryptosporidium-Associated Diarrhea in a High Altitude-Community of Saudi Arabia Detected by Conventional and Molecular Methods
Image Image Image
Fig. 1 Microscopic picture of Cryptosporidium oocysts (red) in a wet mount smear prepared from a fecal specimen concentrated by modified formol-ether method and stained following the modified Zeihl Neelsen procedure (×400 magnification).
Fig. 2 Representative ethidium bromide-stained 1% agarose gel electrophoresis picture showing the PCR test results of 12 samples (Lane 1-12) diagnosed as Cryptosporidium-negative by the LF test. M: GeneRuler™ 100 bp DNA marker; Lanes 1, 3, and 12: PCR products, ≈550 bp each, recovered from Cryptosporidium DNA-positive samples. Lane 2, 4-11: Cryptosporidium DNA-negative samples. Lane 13: PCR-negative control (master mix without template DNA).
Fig. 3 Age-distribution of 4 patients diagnosed as Cryptosporidium-negative by microscopy and proved to be positive by the PCR assay.

Fig. 1

Fig. 2

Fig. 3

Prevalence of Cryptosporidium-Associated Diarrhea in a High Altitude-Community of Saudi Arabia Detected by Conventional and Molecular Methods
Microscopy LF test
Total
P-value Positive
Negative
No. of samples % No. of samples % No. of samples % Positive 17 100 0 0 17 100 P = 0.0000b Negative 1a 0.6 162 99.4 163 100 Total 18 10.0 162 90.0 180 100 Microscopy PCR assay
Total
P-value Positive
Negative
No. of samples % No. of samples % No. of samples % Positive 17 100 0 0 17 100 P = 0.0000b Negative 4a 2.5 159 97.5 163 100 Total 21 11.6 159 88.4 180 100 LF test PCR assay
Total
P value Positive
Negative
No. of samples % No. of samples % No. of samples % Positive 18 100 0 0 18 100 P = 0.0000b Negative 3a 1.9 159 98.1 163 100 Total 21 11.6 159 88.4 180 100 Age groups PCR test results
Total
P-value Positive
Negative
No. of samples % No. of samples % No. of samples % < 5 years 11 22.0 39 78.0 50 100 χ2 = 8.11 5-9 years 5 11.1 40 88.9 45 100 P = 0.004 10-39 years 4 10.0 36 90.0 40 100 40-59 years 0 0 20 100 20 100 ≥ 60 years 1 4.0 24 96.0 25 100 Total 21 11.7 159 88.3 180 100 Gender PCR test results
Total
P value of difference Positive
Negative
No. of samples % No. of samples % No. of samples % Males 12 16.2 62 83.8 74 100 χ2 = 2.5 Females 9 8.5 97 91.5 106 100 P = 0.11 Total 21 11.7 159 88.3 180 100 Diagnostic test Test results
Test diagnostic performance parameters (%)
Positive
Negative
SE SP NPV PPV (No.) (%) (No.) (%) Microscopy 17 9.4 163 90.6 81.0 100 97.6 100 LF test 18 10.0 162 90.0 85.7 100 98.2 100 PCR test 21 11.6 159 88.3 - - - -
Table 1. Concordance (number and percentage) of the microscopy results with the LF test

One sample showed a discordance result (negative by microscopy but positive by the LF test) and proved to be positive by PCR.

Highly significant by Fisher’s exact test.

Table 2. Concordance (number and percentage) of microscopy results with the PCR assay

Samples showed discordance results (these 4 samples were oocyst-negative by microscopy but proved to be Cryptosporidium DNA-positive by PCR).

Highly significant by Fisher’s exact test.

Table 3. Concordance (number and percentage) of the LF test results with the PCR assay

Samples with discordance results (i.e., these 3 samples were negative for Cryptosporidium oocysts by microscopy as well as by the LF test but proved to be positive for DNA by the PCR assay).

Highly significant by Fisher’s exact test.

Table 4. PCR test results of 180 samples distributed by age groups
Table 5. PCR test results (number and percentage) distributed by gender
Table 6. The diagnostic performance of microscopy and the LF test versus the PCR test results

SE, sensitivity; SP, specificity; NPV, negative predictive value; PPV, positive predictive value.

Table 1.

Table 2.

Table 3.

Table 4.

Table 5.

Table 6.

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