Drug resistance-associated mutations in antiretroviral treatment-experienced patients in kuwait

Objectives: To investigate the prevalence of nonpolymorphic resistance-associated mutations [RAM] in HIV-1 patients on first-line antiretroviral therapy in Kuwait

Subjects and Methods: Total RNA was isolated from plasma samples of 42 patients who received a first-line nonnucleoside reverse transcriptase inhibitor [NNRTI]-based regimen. HIV-1 protease and reverse transcriptase genetic regions were then amplified by nested reverse transcription-polymerase chain reaction and directly sequenced. The HIV-1 subtype was identified using the Bayesian phylogenetic method, and RAM were identified using the Stanford University genotypic resistance interpretation algorithm

Results: The HIV-1 viral load at sampling ranged from < 20 to 8.25 × 10[4] copies/ml. CRF01_AE, C, and B were the most predominant HIV-1 subtypes. Nonpolymorphic mutations associated with resistance to antiretroviral drugs were detected in 11 [26.2%] of the 42 patients; 5 [11.9%] patients had mutations associated with a high-level resistance to nucleoside reverse transcriptase inhibitors [NRTI], 4 [9.5%] patients had mutations associated with resistance to NNRTI, 1 [2.4%] patient had mutations associated with resistance to both NRTI and NNRTI, and 1 [2.4%] patient had mutations potentially associated with low-level resistance to both protease inhibitors and NNRTI. All patients with RAM had a detectable plasma HIV-1 RNA level

Conclusion: Our results indicate the development of RAM during an NNRTI-based regimen and highlight the importance of considering other regimens to avoid treatment failure

Phylogenetic analysis of human metapneumovirus detected in hospitalized patients in Kuwait during the years 2009–2011

Human metapneumovirus [hMPV] is an important cause of both upper and lower respiratory tract infections [RTIs] in all age groups. Children, elderly, and immunocompromised individuals are the most affected groups. HMPV infection accounts for 5% of hospitalized patients with respiratory tract infections in Kuwait. It is mostly detected among infants and elderly age groups, and both hMPV genotypes A and B circulate in Kuwait. In this study, the genetic diversity of detected hMPV was evaluated, and a phylogenetic analysis based on partial nucleotide and amino acid sequences of the G gene was performed for hMPV detected among hospitalized patients with RTIs. Our results showed that 62% of hMPV sequences belonged to the A genotype and 38% to the B genotype. A2b and B2 subtypes were detected and circulated during the study period, whereas A1 and B1 subtypes were not detected. Based on nucleotide sequences of the G gene, most of hMPV strains [57%] were clustered with Indian strains, followed by Greek strains [24%] and Canadian strains [14%]. One strain [5%] clustered within the B genotype but had different branches than B1 and B2 branches. Our data showed the co-circulation of hMPV genotypes A2b and B2 in Kuwait with genetic diversity suggestive of evolution through negative selection

prevalence of human Bocavirus, human Coronavirus-NL63, human Metapneumovirus, human Polyomavirus KI and WU in respiratory tract infections in Kuwait

Objective: The aim of this study was to investigate the prevalence of human coronavirus [HCoV]-NL63, human metapneumovirus [hMPV], human bocavirus [Boca], human polyomavirus KI [KIV] and human polyomavirus WU [WUV] in respiratory tract infections [RTI] in Kuwait

Materials and Methods: Respiratory samples from 735 hospitalized patients with RTI from September 2010 to April 2013 were evaluated for the presence of HCoV-NL63, hMPV, Boca, KIV and WUV using molecular assays, polymerase chain reaction [PCR] and reverse-transcription PCR

Results: Of the 735 patients, 285 [38.8%] were diagnosed with viral RTI. The distribution of respiratory viruses was hMPV: 15 [5.3%], Boca: 14 [4.9%], WUV: 10 [3.5%] and KIV: 4 [1.4%]. HCoV-NL63 was not detected in any of the samples

Conclusions: These newly discovered viruses were associated with the development of RTI in Kuwait. The rapid identification of these viral infections could aid in the control of nosocomial transmission, reduce the use of antibiotics and improve treatment and management strategies

Who collaborating centre for acquired immunodeficiency syndrome for the eastern mediterranean regional office, faculty of medicine, Kuwait University, Kuwait

In the early 1980s, the World Health Organization [WHO] designated the Virology Unit of the Faculty of Medicine, Health Sciences Centre, Kuwait University, Kuwait, a collaborating centre for AIDS for the Eastern Mediterranean Regional Office [EMRO], recognizing it to be in compliance with WHO guidelines. In this centre, research integral to the efforts of WHO to combat AIDS is conducted. In addition to annual workshops and symposia, the centre is constantly updating and renewing its facilities and capabilities in keeping with current and latest advances in virology. As an example of the activities of the centre, the HIV-1 RNA viral load in plasma samples of HIV-1 patients is determined by real-time PCR using the AmpliPrep TaqMan HIV-1 test v2.0. HIV-1 drug resistance is determined by sequencing the reverse transcriptase and protease regions on the HIV-1 pol gene, using the TRUGENE HIV-1 Genotyping Assay on the Open-Gene® DNA Sequencing System. HIV-1 subtypes are determined by sequencing the reverse transcriptase and protease regions on the HIV-1 pol gene using the genotyping assays described above. A fundamental program of Kuwait's WHO AIDS collaboration centre is the national project on the surveillance of drug resistance in human deficiency virus in Kuwait, which illustrates how the centre and its activities in Kuwait can serve the EMRO region of WHO

relatively high number of pregnant women in Kuwait remain susceptible to rubella: a need for an alternative vaccination policy

To measure the prevalence of anti-rubella IgG and hepatitis B surface antigen [HBsAg] among pregnant women in Kuwait in order to assess the effectiveness of the current vaccination programs. This retrospective study involved 4,062 pregnant women evaluated in health centers in the Hawalli Province of Kuwait. They were screened for anti-rubella IgG and HBsAg using commercially available assays. The data were obtained from medical laboratory records. The mean age of the pregnant women was 29.2 +/- 5.26 years [range 17-49]. The rubella IgG prevalence among the pregnant women was 88.4% [n = 3,589]; 276 [6.8%] of the pregnant women had no antibody to rubella, and 197 [4.8%] had rubella antibody levels /= 40 years, respectively [p = 0.016]. The prevalence of HBsAg was 0.3%, and it did not vary with age. The prevalence of both anti-rubella IgG and HBsAg among pregnant women in Kuwait was relatively high. However, about 11.6% of pregnant women in Kuwait remain susceptible to rubella infection and hence congenital infection and fetal malformation

Hepatitis C virus infection in a population with high incidence of type 2 diabetes: Impact on diabetes complications

A growing number of reports suggest a connection between hepatitis C virus [HCV] infection and type 2 diabetes [T2D]. However, the association of HCV infection with diabetes-related complications has not yet been clarified. The aim of this study was to determine the prevalence of HCV infection in T2D-patients in Kuwait which has a high incidence of type 2 diabetes, and to investigate the association between HCV viremia and diabetes-related complications. A total of 438 patients with T2D [325 Kuwaitis and 113 Egyptians], and 440 control subjects, were enrolled for this study. HCV infection was assessed by testing for serum HCV-specific antibodies, and by detection of HCV RNA. HCV viral load and hemoglobin A1c [HbA1c] levels were assessed in patients with and without diabetes complications. Thirty one [7%] out of 438 T2D-patients had evidence of HCV infection compared to 4 [1%] out of 440 control adults [p < 0.0001]. The prevalence of HCV infection in Kuwaiti and Egyptian T2D-patients was 3% and 18%, respectively. Most of the HCV sequences detected in T2D patients and control subjects were of genotype 4. The HbA1c levels in T2D-patients with HCV viremia were significantly higher than those in HCV-negative patients. HCV viremia, female sex, age, family history of diabetes were found to be independent risk factors for diabetes complications. The results suggest that T2D-patients in Kuwait have higher prevalence of HCV infection than controls, and that HCV viremia is associated with diabetes-related complications

Detection and monitoring of cytomegalovirus infection in renal transplant patients by quantitative reAl time PCR

To establish a sensitive and specific real time PCR for quantitation of cytomegalovirus [CMV] DNA in clinical specimens. In a prospective study, CMV DNA was quantified in blood samples of 255 kidney recipients with and without CMV-related symptoms between the years 2000 and 2005 in Kuwait. In a selected group of patients, the effect of anti-CMV chemotherapy was monitored by quantitative real time PCR [qRT-PCR]. The established qRT-PCR assay had a sensitivity to detect 30 CMV DNA copies. CMV DNA was detected in 54/255 [24%] patients; of these, 17 [31.5%] were asymptomatic, and 37 patients [68.5%] had symptomatic CMV infection. Sequential blood specimens were collected from all CMV-positive patients and tested by CMV pp65 antigenemia and qRT-PCR assays. There was a moderate positive correlation between the two assays [Pearson's correlation=0.52]. The median CMV viral load measured by qRT-PCR was higher in symptomatic [6.5 +/- 10.4 copies/ml] than in asymptomatic [185 copies/ml] patients [p=0.001]. The estimated cut-off value of CMV DNA for CMV symptoms/disease was 6 800 copies/ml of blood. Testing of sequential samples from patients treated with symptomatic CMV infection showed that the viral load was significantly reduced after 3 weeks of anti-CMV chemotherapy [p=0.001]. The reported qRT-PCR is a sensitive method for quantitation of CMV DNA in the blood of kidney recipients and can be useful in monitoring the efficacy of anti-CMV therapy

rapid and sensitive method for the diagnosis of the "avian flu" H5 sub-type of influenza A virus

Avian influenza, an infectious disease of birds, is caused by type A strain of influenza virus. Within the first 3 months of 2006. 46 human cases of avian influenza in humans worldwide were found where of 31 have died with a mortality rate of about 67%. Therefore, the need for a rapid and sensitive method to diagnose the infection, subtype, and quantify the virus is of paramount importance in the management of this pandemic. Virology unit, Kuwait University, Mubarak hospital in Kuwait. 80 patients with lower respiratory tract infections. We have developed a sensitive and specific multiplex RT-PCR capable of detecting common nine respiratory viruses causing acute respiratory disease in Kuwait. Those found positive for influenza A virus, were then subjected to a quantitative Real-Time PCR to sub-type the virus as H5 and to measure the amount of virus in respiratory samples. A 189 bp fragment of the influenza virus A H5 gene was amplified with specific primers and quantitatively detected with probes supplied in the LightMix [TIB MOLBIOL, Berlin]. The supplied standard dilutions of influenza A H5 virus cDNA ranging from 101 to 106 copies/reaction allows the absolute quantification of the viral cDNA in the unknown samples. A standard curve using standard increasing dilutions of influenza A H5 ranging from 101 to 106 cDNA copies /ml was plotted to quantitate influenza A virus of the H5 subtype. The minimum detection limit was 10 copies/ml. Specificity of the primer/probe mixture was tested on samples containing rhino-, corona-, RSV, measles and mumps viruses. Out of the 10 of 80 [12.5%] patients with acute respiratory disease who were influenza virus A positive, none was found to be of the H5 sub-type of influenza A virus so far in Kuwait. The combination of the multiplex RT-PCR developed in our laboratory and the real time PCR provides a fast easy, specific, and accurate system for the detection of influenza A H 5 subtype