1.Detection of α-thalassaemia in neonates on cord blood and dried blood spot samples by capillary electrophoresis
Hafiza Alauddin ; Mustafa Langa ; Malisa Mohd Yusoff ; Raja Zahratul Azma Raja Sabudin ; Mpath, Azlin Ithnin ; Noor Hamidah Hussin
The Malaysian Journal of Pathology 2017;39(1):17-23
Haemoglobin Bart’s (Hb Bart’s) level is associated with α-thalassaemia traits in neonates,
enabling early diagnosis of α-thalassaemia. The study aimed to detect and quantify the Hb Bart’s
using Cord Blood (CB) and CE Neonat Fast Hb (NF) progammes on fresh and dried blood spot
(DBS) specimen respectively by capillary electrophoresis (CE). Methods: Capillarys Hemoglobin
(E) Kit (for CB) and Capillarys Neonat Hb Kit (for NF) were used to detect and quantify Hb Bart’s
by CE in fresh cord blood and dried blood spot (DBS) specimens respectively. High performance
liquid chromatography (HPLC) using the β-Thal Short Programme was also performed concurrently
with CE analysis. Confirmation was obtained by multiplex ARMS Gap PCR. Results: This study
was performed on 600 neonates. 32/600 (5.3%) samples showed presence of Hb Bart’s peak using
the NF programme while 33/600 (5.5%) were positive with CB programme and HPLC methods.
The range of Hb Bart’s using NF programme and CB programme were (0.5–4.1%) and (0.5-7.1%),
respectively. Molecular analysis confirmed all positive samples possessed α-thalassaemia genetic
mutations, with 23/33 cases being αα/--SEA, four -α3.7/-α3.7, two αα/-α3.7 and three αα/ααCS. Fifty Hb
Bart’s negative samples were randomly tested for α-genotypes, three were also found to be positive
for α-globin gene mutations. Thus, resulting in sensitivity of 91.7% and 88.9% and specificity of
100% for the Capillarys Cord Blood programme and Capillarys Neonat Fast programme respectively.
Conclusion: Both CE programmes using fresh or dried cord blood were useful as a screening tool
for α-thalassaemia in newborns. All methods show the same specificity (100%) with variable, but
acceptable sensitivities in the detection of Hb Bart.
2.In Vitro Antifungal Activities against Moulds Isolated from Dermatological Specimens
Tzar Mohd Nizam ; Rabiatul Adawiyah AG. Binting ; Shafika Mohd Saari ; Thivyananthini Vijaya Kumar ; Marianayati Muhammad ; Hartini Satim ; Hamidah Yusoff ; Jacinta Santhanam
Malaysian Journal of Medical Sciences 2016;23(3):32-39
Background: This study aimed to determine the minimum inhibitory concentrations (MICs) of various antifungal agents against moulds isolated from dermatological specimens. Methods: We identified 29 moulds from dermatological specimens between October 2012 and March 2013 by conventional methods. We performed antifungal susceptibility testing on six antifungal agents, amphotericin B, clotrimazole, itraconazole, ketoconazole, miconazole and terbinafine, according to the Clinical and Laboratory Standards Institute guidelines contained in the M38-A2 document. Results: Most antifungal agents were active against the dermatophytes, except for terbinafine against Trichophyton rubrum (geometric mean MIC, MICGM 3.17 µg/mL). The dematiaceous moulds were relatively susceptible to amphotericin B and azoles (MICGM 0.17-0.34 µg/mL), but not to terbinafine (MICGM 3.62 µg/mL). Septate hyaline moulds showed variable results between the relatively more susceptible Aspergillus spp. (MICGM 0.25-4 µg/mL) and the more resistant Fusarium spp. (MICGM 5.66-32 µg/mL). The zygomycetes were susceptible to amphotericin B (MICGM 0.5 µg/mL) and clotrimazole (MICGM 0.08 µg/mL), but not to other azoles (MICGM 2.52-4 µg/mL). Conclusion: Amphotericin B and clotrimazole were the most effective antifungal agents against all moulds excepting Fusarium spp., while terbinafine was useful against dermatophytes (except T. rubrum) and Aspergillus spp. However, a larger study is required to draw more solid conclusions.
3.HbA2 levels in normal, B-thalassaemia and haemoglobin E carriers by capillary electrophoresis
Hafiza Alauddin ; Malisa Mohd Yusoff ; RD Aidifitrina Khirotdin ; Azlin Hanim ; Raja Zahratul Azma ; Matthew Chong Kwok Thong ; Irwan Mohamad Ali ; Yeoh Zi-Ning ; Lailyvia Mohd Ishak ; Nur Rabiatul Adawiah Mohd Radzi ; Noor Hamidah Hussin
The Malaysian Journal of Pathology 2012;34(2):161-164
Objective: The capillary electrophoresis (CE) is a new system that utilizes the principle of electrokinetic
separation of molecules in eight electrolyte buffer-fi lled silica capillaries. In this study, we established
the normal ranges of haemoglobin A2 (HbA2) and haemoglobin F (HbF) levels for normal individuals
using this system and also the HbA2 level in β thalassaemia and haemoglobin E (HbE) individuals.
Materials and Methods: 154 samples from normal individuals, 218 samples from β thalassaemia
heterozygotes and 91 samples from HbE heterozygotes were subjected to high performance liquid
chromatography (HPLC) and CE analysis. Results: The normal ranges for HbA2 and HbF by CE
were 2.75% (SD 0.26%) and 0.03% (SD 0.24%) respectively, which were signifi cantly lower than
that of HPLC 2.88% (SD 0.25%) and 0.58% (SD 0.61%) (p <0.001). The HbA2 level for HbE
heterozygotes was 3.58% (SD 0.44%), which was signifi cantly higher than normal (p <0.001) but
lower than that of β-thalassaemia heterozygotes (p<0.001) and the true HbE level was 24.28% (SD
3.38%). Conclusion: The CE system provided a fully automated and high throughput system for
haemoglobin analysis. We established the normal ranges for HbA2 and HbF levels by CE. We also
determined the ranges for HbA2 in beta thalassaemia and HbE heterozygotes using this system.
4.HbA2 levels in normal, beta-thalassaemia and haemoglobin E carriers by capillary electrophoresis.
Hafiza, Alauddin ; Malisa, Mohd Yusoff ; Khirotdin, R D Aidifitrina ; Azlin, Ithnin ; Azma, Zahratul ; Thong, Matthew Chong Kwok ; Ali, Irwan Mohamad ; Yeoh, Zi-Ning ; Mohd Ishak, Lailyvia ; Mohd Radzi, Nur Rabiatuladawiah ; Hussin, Noor Hamidah
The Malaysian Journal of Pathology 2012;34(2):161-4
The capillary electrophoresis (CE) is a new system that utilizes the principle of electrokinetic separation of molecules in eight electrolyte buffer-filled silica capillaries. In this study, we established the normal ranges of haemoglobin A2 (HbA2) and haemoglobin F (HbF) levels for normal individuals using this system and also the HbA2 level in beta thalassaemia and haemoglobin E (HbE) individuals.
5.The crucial role of molecular testing to facilitate the diagnosis of pneumocystis pneumonia during pregnancy
Chuan Hun DING ; Hamidah YUSOFF ; Najihan Abdul Samat Muttaqillah ; Yee Loong TANG ; Toh Leong TAN ; Petrick PERIYASAM ; Andrea Yu-Lin BAN
The Malaysian Journal of Pathology 2018;40(1):67-72
Pneumocystis pneumonia is an important human immunodeficiency virus (HIV)-associated opportunistic infection, and especially so in pregnant HIV-positive patients. We report a case of a 40-year-old woman in her first trimester of pregnancy who initially presented with acute gastroenteritis symptoms but due to a history of high-risk behaviour and the observation of oral thrush, she was worked up for HIV infection. Her retroviral status was positive and her CD4+ T cell count was only 8 cells/mL. She was also worked up for pneumocystis pneumonia due to the presence of mild resting tachypnoea and a notable drop in oxygen saturation (from 100% to 88%) following brief ambulation. Her chest radiograph revealed bilaterally symmetrical lower zone reticular opacities and Giemsa staining of her bronchoalveolar lavage (BAL) was negative for Pneumocystis jirovecii cysts. However, real-time P. jirovecii polymerase chain reaction (PCR) testing on the same BAL specimen revealed the presence of the organism. A course of oral co-trimoxazole plus prednisolone was commenced and her clinical condition improved.