Detection of asymptomatic Leishmania donovani in healthy voluntary blood donors.

OBJECTIVE: To check incidence of Asymptomatic Leishmania donovani reporting to Armed Forces Institute of Transfusion Rawalpindi. MATERIAL AND METHODS: Two thousand (n = 2000) consecutive healthy voluntary blood donors were tested for 18 s rRNA by Real time Polymerase chain reaction. One thousand (n = 1000) subject's permanent resident of Azad Kashmir along with a thousand (n = 1000) healthy voluntary blood donors from rest of Pakistan were included. The study was carried out over a period of three months Jun - Aug 2020. RESULTS: Total of 2000 blood donors were enrolled in the study, all were males with age ranging from 16 to 60 years. Stratification based on residence, 1000 (50 %) resided in the Azad kashmir, 349 (17.45 %) were from Islamabad and Rawalpindi, 541 (27.05 %) from Punjab mainly residing in Lahore and Multan, 110 (5.5 %) were from other cities of Pakistan. Grouping on the basis of age, 55.25 % (n = 1105) of the donors were 16-25 years old, 19.45 % (n = 389) were in age range of 26-40 years old, 15.55 % (n = 311) were 41-50 years old and 9.75 % (n = 195) 51-60 years old. No donor was diagnosed as an asymptomatic carrier. CONCLUSION: Screening of blood donors for Leishmania donovani is not recommended.

Noninvasive prenatal testing of beta-thalassemia for common Pakistani mutations: a comparative study using cell-free fetal DNA from maternal plasma and chorionic villus sampling.

BACKGROUND: The discovery of circulating cell-free fetal DNA (cff-DNA) in maternal plasma has inspired the noninvasive prenatal testing (NIPT) approaches for various genetic fetal screening including rhesus D typing, sex determination, aneuploidies, and single-gene disorders. OBJECTIVE: Noninvasive determination of paternally inherited beta-thalassemia mutations in maternal total cell-free DNA (cf-DNA) by using allele-specific amplification refractory mutation system (ARMS) real-time PCR (RT-PCR) in concordance with the conventional invasive method. METHODS: An observational study was conducted at the Armed Forces Institute of Blood Transfusion in collaboration with the genetics resource center from March 2021 to August 2021. A total number of 26 couples were selected having a history of previously affected children with beta-thalassemia. A routine chorionic villus sampling (CVS) invasive procedure was carried out, and the mutation analysis was done using conventional PCR. To assess NIPT, a total cf-DNA was also extracted from maternal plasma and analyzed using allele-specific ARMS RT-PCR. RESULTS: Based on conventional PCR testing, 13 of 26 couples were found having beta-thalassemia carriers with homozygous mutation, and 13 couples were carriers with heterozygous mutations. Further to assess NIPT, the cf-DNA of 13 pregnant females among the couples with different mutational patterns was analyzed by allele-specific ARMS RT-PCR to detect paternally inherited mutations. In comparison with conventional PCR, 11 cases (84.6%) were matched successfully, while two cases (15.4%) had no concordance with conventional invasive prenatal testing (IPT). CONCLUSION: NIPT using maternal cf-DNA by allele-specific ARMS RT-PCR can be feasible to screen paternal inherited mutant alleles to rule out pregnant women from invasive procedures where the test would be negative for paternal inheritance. However, a low amount of fetal DNA in maternal plasma is a limiting factor and required further improvement to enrich fetal cf-DNA for complete concordance with conventional IPT.