Sains Malaysiana 47(9)(2018): 2099–2104

http://dx.doi.org/10.17576/jsm-2018-4709-18

Comparison of Direct Examination and In Vitro Culture for the Detection of Blastocystis sp. in Orang Asli Stool Samples

(Perbandingan Pemeriksaan Langsung dan Kultur In Vitro untuk Pengesanan Blastocystis sp. dalam Spesimen Najis Orang Asli)

SAMSEH ABDULLAH NORADILAH¹, NORHAYATI MOKTAR², II LI LEE³, FATMAH MD SALLEH⁴ & TENGKU SHAHRUL ANUAR^5,6*

¹Department of Medical Sciences II, Faculty of Medicine and Health Sciences, Universiti Sains Islam Malaysia,55100, Pandan Indah, Kuala Lumpur, Federal Territory, Malaysia

²Department of Pre-Clinical Sciences, Faculty of Medicine and Health Sciences, Universiti Tunku Abdul Rahman, Sungai Long Campus, 43000 Kajang, Selangor Darul Ehsan, Malaysia

³Kulliyyah of Medicine and Health Sciences, Sultan Abdul Halim Mu’adzam Shah International Islamic University, 09300 Kuala Ketil, Kedah Darul Aman, Malaysia

⁴Department of Parasitology and Medical Entomology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

⁵Centre of Medical Laboratory Technology, Faculty of Health Sciences, Universiti Teknologi MARA, Puncak Alam Campus, 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia

⁶Integrative Pharmacogenomics Institute, Universiti Teknologi MARA, Puncak Alam Campus, 42300 Bandar Puncak Alam, Selangor Darul Ehsan, Malaysia

Diserahkan: 8 Februari 2018/Diterima: 14 Mei 2018

ABSTRACT

Microscopy-based technique has been widely used in the detection of Blastocystis sp. This study was conducted to compare the techniques used for screening of Blastocystis sp., namely in vitro cultivation of stool specimens in Jones’ medium (IVC) followed by Wheatley Trichrome staining and direct examination of stool samples preserved with polyvinyl alcohol (PVA) followed by Wheatley Trichrome staining with single-round polymerase chain reaction (PCR) as the reference technique. The study was performed on 466 stool samples obtained from the aboriginal community in Pahang, Malaysia. IVC showed higher detection rate of Blastocystis sp. (35.6%) than PVA (20.0%). Single-round PCR detected Blastocystis sp. in 41.0% of the stool specimens. The sensitivity and specificity of PVA and IVC in comparison to the reference technique were 75.3% (95% CI: 65.2-83.6) and 68.5% (CI: 63.7-73.3) and 88.6% (CI: 82.7-93.0) and 86.3% (CI: 81.9-90.0), respectively. The agreement between the reference technique and PVA showed statistically significant fair agreement by Cohen Kappa statistics of (K=0.318, p<0.001), meanwhile statistically significant substantial agreement was observed between PCR and IVC by Cohen Kappa (K=0.727, p<0.001). Therefore, in vitro cultivation in Jones’ medium followed by Wheatley Trichrome staining of stool specimens should be used as a screening technique in the detection of Blastocystis sp. infections.

Keywords: Blastocystis detection; in vitro cultivation; microscopy; polymerase chain reaction; polyvinyl alcohol

ABSTRAK

Teknik berasaskan mikroskopi telah digunakan secara meluas dalam pengesanan Blastocystis sp. Kajian ini dijalankan untuk membandingkan dua teknik saringan; kultur secara in vitro spesimen najis dalam media Jones’ (IVC) diikuti dengan pewarnaan Trikrom Wheatley dan pemeriksaan langsung sampel najis yang diawet dengan alkohol polivinil (PVA) diikuti dengan pewarnaan Trikrom Wheatley dengan satu pusingan tindak balas berantai polimerase (PCR) sebagai teknik rujukan. Kajian ini dijalankan terhadap 466 sampel najis yang diperoleh daripada komuniti Orang Asli di Pahang, Malaysia. Teknik IVC menunjukkan kadar pengesanan yang lebih tinggi (35.6%) berbanding PVA (20.0%). Kesensitifan dan kekhususan  PVA dan IVC berbanding teknik rujukan masing-masing adalah 75.3% (95% CI: 65.2-83.6) dan 68.5% (CI: 63.7-73.3) dan 88.6% (CI: 82.7-93.0) dan 86.3% (CI: 81.9-90.0). Darjah persetujuan antara teknik rujukan dan PVA menunjukkan persetujuan sederhana menggunakan analisis Cohen Kappa (K=0.318, p<0.001), manakala persetujuan yang baik ditunjukkan antara PCR dan IVC menggunakan analisis yang sama (K=0.727, p<0.001). Oleh yang demikian, teknik kultur secara in vitro dalam media Jones’ diikuti pewarnaan Trikrom Wheatley untuk spesimen najis harus digunakan sebagai teknik saringan dalam pengesanan jangkitan Blastocystis sp..

Kata kunci: Alkohol polivinil; kultur secara in vitro; mikroskopi; pengesanan Blastocystis; tindak balas berantai polymerase

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*Pengarang untuk surat-menyurat; email: noradilah82@gmail.com