Drug resistance mutations among virological failure HIV-1 infected patients in Malaysia.

The determination of HIV drug resistance mutations (DRMs) towards antiretroviral (ARV) drugs among HIV-1 treated patients with virological failure is crucial for further management of the patient. This study aimed to assess the most common genomic mutation and to analyse subtypes among the HIV-1 patients with viral load level > 1,000 copies/mL. A total of 101 virological failure HIV-1 patients from four different regions of Peninsular Malaysia with a viral load measurement facility were included in the study. Majority of patients (89.1%) have at least 1 mutation associated with clinical resistance to either protease inhibitors (PIs), nucleoside reverse transcriptase inhibitors (NRTIs) or nonnucleoside reverse transcriptase inhibitors (NNRTIs). Major resistance mutations among the patients towards NRTIs and NNRTIs were 70.3% and 18.8%, respectively. The most common mutation for NRTIs was M184V while K103N mutation was detected in the majority of patients who were treated with NNRTIs. The most commonly observed mutations for major PI and minor PI seen among the study population were V82A/T and L10V, respectively. In HIV-1 subtype analysis, CRF33_01B was the most predominant HIV-1 subtype in this study group. The vast detection of DRMs in this study emphasized the importance of genotypic resistance test in the management of HIV patients as DRMs can alter patient's susceptibility towards ARV drugs. Further study on larger number of samples is essential for the development of a database on HIV-1 DRMs among patients that experience virological failure in Malaysia.

Dengue serotype surveillance among patients admitted for dengue in two major hospitals in Selangor, Malaysia, 2010-2011.

Dengue serotype surveillance is important as any changes in serotype distribution may result in an outbreak or increase in severe dengue cases. This study aimed to determine circulating dengue serotypes in two hospitals in Selangor. Serum samples were collected from patients admitted for dengue at these two major public hospitals i.e. Hospital Sungai Buloh (HSB) and Hospital Tunku Ampuan Rahimah (HTAR) between November 2010 and August 2011 and subjected to real-time RT-PCR using SYBR® Green. All four dengue serotypes were detected in samples from both hospitals. The predominating serotype was dengue 1 in samples from both hospitals (HSB, DENV-1; 25.53 % and HTAR, DENV-1; 32.1 %).

Combined detection and genotyping of Chikungunya virus by a specific reverse transcription-polymerase chain reaction.

A reverse transcription-polymerase chain reaction (RT-PCR) was developed for the detection of Chikungunya virus infection. Based on the nonstructural protein 1 (nsP1) and glycoprotein E1 (E1) genes of Chikungunya, two primer sets were designed. Total RNA were extracted from the cell culture fluid of Aedes albopictus C6/36 cells inoculated with the S27 prototype virus, isolated in Tanzania in 1953, and the Malaysian strains (MALh0198, MALh0298, and MALh0398), isolated in Malaysia in 1998. For both sets of RNA samples, the expected 354- and 294-base pair (bp) cDNA fragments were amplified effectively from the nsP1 and E1 genes, respectively. Phylogenetic analysis was conducted for the Malaysian strain and other virus strains isolated from different regions in the world endemic for Chikungunya, using partial E1 gene sequence data. The Malaysian strains isolated during the epidemics of 1998 fell into a cluster with other members of the Asian genotype.

Type-3 dengue viruses responsible for the dengue epidemic in Malaysia during 1993-1994.

To characterize the dengue epidemic that recently occurred in Malaysia, we sequenced cDNAs from nine 1993-1994 dengue virus type-3 (DEN-3) isolates in Malaysia (DEN-3 was the most common type in Malaysia during this period). Nucleic acid sequences (720 nucleotides in length) from the nine isolates, encompassing the precursor of membrane protein (preM) and membrane (M) protein genes and part of the envelope (E) protein gene were aligned with various reference DEN-3 sequences to generate a neighbor-joining phylogenetic tree. According to the constructed tree, the nine Malaysian isolates were grouped into subtype II, which comprises Thai isolates from 1962 to 1987. Five earlier DEN-3 virus Malaysian isolates from 1974 to 1981 belonged to subtype I. The present data indicate that the recent dengue epidemic in Malaysia was due to the introduction of DEN-3 viruses previously endemic to Thailand.

A four year review of acute viral hepatitis cases in the east coast of peninsular Malaysia (1994-1997).

A total of 1,157 sera from jaundiced patients with clinical and biochemical evidence of liver disease received from government hospital in Kelantan and Terengganu, during the period from 1994 to 1997, were investigated to determine the cause. Hepatitis A virus was found to be the main cause in 26.1% (24/92) of symptomatic clinical hepatitis cases in 1994, 47.8% (63/132) in 1995, 66.4% (613/923) in 1996 and 20% (2/10) in 1997. Sera received in 1996 were also tested for hepatitis B, hepatitis C, hepatitis D and hepatitis E. 1.4% (13/923) anti-bodies were found to be positive for HBc IgM indicating recent HBV infection, 5.4% (50/923) for total HCV Ab, 0.9% (8/923) for total HDV Ab and 0.4% (4/923) for anti-HEV IgM. This study shows that HAV is still a major problem in Kelantan and Terengganu, and there is a need to identify effective strategies for prevention and control in these two states.

On-the-job training through follow-up visits to improve the quality of family planning services.

As part of a broader set of activities to strengthen family planning training and improve the quality of family planning services in Turkey, follow-up visits were performed at different family planning sites across the country in order to conduct on-the-job training. The objective of on-the-job training was to refresh and improve family planning counselling skills for all methods as well as to refresh and improve intrauterine device insertion/removal skills and also some determinants of quality care. It was also aimed at transferring up-to-date information to family planning practitioners, identifying frequently encountered problems and helping with solution approaches for problems both at the individual and programmatic levels. The results of the follow-up visits reflect issues about both the staff and the clinical facility itself in terms of conforming with the standards of the 'National Family Planning Guidelines' set forth by the Ministry of Health. The follow-up team consisted of nine members who were specially trained. They represented different sectors such as a non-governmental organization, a medical school and the Ministry of Health. The follow-up team performed 90 visits to 16 clinics in 11 provinces between 1995 and 1998. Methods used were structured observations via standard checklists, meetings with the clinic staff, self-assessment, role plays, demonstration, coaching and the provision of feedback. During this period, a total of 130 health professionals working in 16 clinics were trained on-the-job. A significant improvement was observed in the performance of the family planning practitioners and the quality of care provided in clinics. While none of the service providers were found to have a standard skill level in general counselling during the first visit, at the end of the fifth visit all were capable of providing counselling services according to the national standards. Intrauterine device insertion skills were high at the beginning of the visits, and 16 of the 17 observed service providers (94%) were assessed as conforming to the standards. At the fifth visit, all of the 42 service providers (100%) were found to be adequate. At the facility level, all 16 clinics established separate counselling rooms in the follow-up period. Additionally, the number of clinics conforming to infection prevention standards increased from two clinics in 15 at the first visit to all 16 clinics at the fifth visit. This study showed that the ultimate success of family planning programs depend on structured and well-supervised on-the-job training through follow-up visits to the sites.

Detection of dengue virus from field Aedes aegypti and Aedes albopictus adults and larvae.

Mosquito adults and larvae were collected from dengue high risk areas and transported to the laboratory for identification. Identified mosquitos were pooled according to the species, date and locality and stored at -70 degrees C. A total of 1,385 pools of Aedes albopictus and 267 pools of Ae. Aegypti were collected from major towns in 12 states in Peninsular Malaysia. Virus isolation was carried out using cell culture (C6/36 clone) of Ae. albopictus and detection of dengue virus by the peroxidase anti-peroxidase staining. All positive isolations were further re-confirmed by the reverse transcriptase-polymerase chain reaction (RT-PCR). Most of the pools were negative with PAP staining and RT-PCR. However, 11 mosquito pools were positive with PAP staining. On the other hand, samples from Terengganu, Pulau Pinang and Johor were positive using both methods.