Minipool Testing Research Articles

Effectiveness Of Pcr Pool Testing For Screening Of Covid-19 Infection In Pakistan.

We tested the utility of mini-pool PCR testing for the rational use of PCR consumables in screening for CoViD-19. After pilot experiments, 3-samples pool size was selected. One step RT-PCR was performed. The samples in the mini-pool having COVID gene amplification were tested individually. 1548 samples tested in 516 mini-pools resulted 396 mini-pools as negative and 120 as positive. Upon individual testing, 110 samples tested positive and 9 were inconclusive. 876 PCR reactions were performed to test 1548 samples, saving 43% PCR reagents. Centres with low prevalence resulted in most saving on reagents (50%), while centres with high prevalence resulted in more test reactions. Testing of individual samples resulted in delays in reporting. Pooling can increase lab capacity, however, pooling delays results and cause degradation of samples.

Blood donor screening in the Netherlands: Universal anti-HBc screening in combination with HBV nucleic acid amplification testing may allow discontinuation of hepatitis B virus antigen testing.

In the Netherlands, blood donor screening for hepatitis B virus (HBV) consists of HBsAg screening since the 1970s, HBV DNA minipool testing (MP-NAT) since 2008, and anti-HBc screening since 2011. Anti-HBc reactivity causes deferral only if anti-HBs titers are <200 IU/mL, or when anti-HBc was acquired during follow-up. Over 5.5 million donations from 582,459 Dutch donors were screened for HBV DNA, HBsAg, anti-HBc, and, if anti-HBc positive, also for anti-HBs. The added value, expressed as the yield of (potentially) infectious and/or recent HBV infections versus unnecessary donor loss, was evaluated for each of the three HBV screening tests. HBV donor screening identified 89 HBV-infected donors with at least two reactive HBV markers (MP-NAT, HBsAg and/or anti-HBc). Single HBV-marker yield was: 5 MP-NAT-only, 0 HBsAg-only, and 20 anti-HBc-only donors. In addition, anti-HBc screening yielded 1,067 potentially infectious donors at risk for occult HBV infection (OBI). In total, 4,126 (0.71%) donors were anti-HBc-reactive at first-time screening, and 1,098 (0.19%) seroconverted during follow-up. Anti-HBc-related donor loss was limited to 2,627 (0.45%) donors using anti-HBs titers and two-strike programs. Donor loss due to MP-NAT and HBsAg screening was extremely low: 0 and 128 donors, respectively. HBV donor screening could be limited to MP-NAT and anti-HBc screening. MP-NAT and anti-HBc improved blood safety by intercepting infectious donations from donors with recent infection or OBI, while HBsAg did not. Unnecessary donor loss related to anti-HBc screening is substantial but does not endanger the continuity of the blood supply.

Donation testing and transfusion transmissible infections

Donation testing and transfusion transmissible infections

No evidence of SARS-CoV-2 RNA among blood donors: A multicenter study in Hubei, China.

BackgroundSevere acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) RNA could be detected in the blood of infected cases. From February 9, all blood establishments in Hubei province, China, implemented nucleic acid testing (NAT) for SARS‐CoV‐2 RNA among blood donors to ensure blood safety.Study Design and MethodsNucleic acid test screening individually (ID) or by minipool (MP) testing was performed according to the manufacturerʼs instructions. Inactivated culture supernatant of SARS‐CoV‐2–infected Vero cells was quantified by droplet digital polymerase chain reaction (ddPCR) and series diluted with negative plasma to evaluate the assayʼs performance.ResultsThe limit of detection of the kit for MP testing was 62.94 and 33.14 copies/mL for N and ORF1ab region, respectively. ID testing could achieve 3.87 and 4.85 copies/mL for two regions using 1600 μL of plasma. Coefficients of variations of two different concentrations of reference samples were all less than 5% in MP testing. As of April 30, 2020, a total of 98,342 blood donations including 87,095 whole blood donations and 11,247 platelet donations were tested by ID or MP testing, and no RNAemia was found. In addition, Hubei province suffered precipitously decreased blood supply, especially in February: 86% reduction compared with the same period of 2019.ConclusionNucleic acid test screening of SARS‐CoV‐2 on blood donations is suitable in blood establishments using the commercial real‐time PCR detection kit based on available instruments. The negative result indicated that SARS‐CoV‐2 appears to be no direct threat to blood safety but raises some serious issues for general blood supply.

First Zika-positive donations in the continental United States.

Zika virus (ZIKV) has spread in the Americas, including parts of the southern United States, and infection can be associated with serious complications, including congenital brain abnormalities. Probable transfusion transmission of ZIKV has been documented in Brazil. Preemptive testing of blood donations for ZIKV RNA was implemented in southern US states at risk of local transmission using a test approved under a Food and Drug Administration (FDA) investigational new drug application, cobas Zika. Screening was expanded after issuance of an updated FDA guidance. Donations reactive on initial screening were further tested by nucleic acid and antibody tests to determine the donor status. Of 358,786 donations from US states screened by individual donation testing, 23 were initially reactive on cobas Zika. Fourteen of these represented probable ZIKV infection based on reactivity on additional nucleic acid testing or anti-Zika immunoglobulin M. Ten of the 14 donors reported travel to an identified ZIKV-active area within 90 days before donation (median time from end of travel to donation, 25 days; range, 6-71 days). Three donors with travel history also had a potential sexual exposure. Only seven of the 14 donations with probable ZIKV infection were detectable upon 1:6 dilution to simulate minipool testing. The estimated specificity of the cobas Zika test was 99.997%. Screening of donations for ZIKV RNA can interdict ZIKV-infected donors. Donor risk factors include travel more than 4 weeks before donation and sexual exposure. Minipool screening would have detected only 50% of the RNA-positive donations.

Prevalence and Trends of Common Transfusion Transmitted Infections Using Serological and Nucleic Acid Markers in Saudi Blood Donors

Background: Epidemiologic studies on the prevalence of transfusion-transmitted infections (TTIs) in Saudi Arabia (SA) regions are limited. This study investigated prevalence of HBV, HCV, and HIV using both serological and nucleic acid testing (NAT) methods to determine temporal and geographic trends among blood donors in Makkah. Our secondary objective was to realize the most suitable NAT format, without compromising sensitivity of NAT results by using individualized or mini-pool testing. Methods: Serologic and NAT screening records of 22,963 blood donors from January 2011 to December 2014 were evaluated for HBsAg, Anti-HBc, Anti-HCV, Anti-HIV, HBV-DNA, HCV-RNA, and HIV-RNA. Prevalence rates were calculated for TTIs per hundred donations and additional analysis was conducted to examine donor profiles associated with positive serologic and NAT results. Known viral loads (<20 IU/ml for each HBV and HCV and <50 copies/ml HIV) diluted in negative plasma were evaluated by NAT screening. Results: Overall serological prevalence of HBs-Ag, anti-HBc, anti-HCV, and anti-HIV were 0.7, 6.7, 0.44, and 0.07%, while molecular HBV-DNA, HCV-RNA, and HIV-RNA were 0.72, 0.05, and 0.03% respectively. There was a gradual decline in percentage of infected donor blood based on combined serological and/or NAT screening from 8.3% in 2011 to 6.8% in 2014 with an overall 7.4% (n=1,689) TTI-infected. Prevalence of HBV, HCV and HIV was unevenly distributed among different regions in SA. Analysis of donor serologic and molecular profiles revealed solitary anti-HBc- positive was the highest (6%) donor profile followed by anti-HBc-positive/HbsAg-positive/HBVDNA positive donor profile at 0.6%, and solitary anti-HCV at 0.4%. Simulation of mini-pool NAT format by dilution of known viral loads at 1:6, resulted in 70% reduction in HBV detection, 50% for HCV, and 40% for HIV. Conclusions: This is the first study to provide current data collectively comparing prevalence and trends of HBV, HCV, and HIV serologic and nucleic acid markers amongst Saudi blood donors. Makkah boasts one of the lowest TTIs prevalence in SA and to surrounding countries. The majority of seropositive and NAT-reactive blood donors are in a state of acute, chronic or resolved HBV infection. Individual donor NAT is the ideal methodology that should be applied in SA where diluted samples could compromise clinical sensitivity and blood safety.

Sensibilidad del equipo Cobas AmpliScreen™ HIV-1 Test, v1.5, para la detección de HIV-1

The introduction of nucleic acid amplification techniques (NAT) in blood banks was intended to reduce the residual risk of transfusion-transmitted infections. Co-circulation of a great diversity of HIV-1 variants in Argentina portrays the need to assess the sensitivity of serological and molecular assays available for their detection. In this study, we evaluated the sensitivity of the COBAS AmpliScreen™ HIV-1 Test, version 1.5 (Roche) for the detection of HIV-1 RNA in plasma samples of infected individuals from Argentina. The results of this study reveal that this technique has high sensitivity for the detection of HIV-1 RNA under assay conditions: using mini-pool testing, pools ≥ 50 RNA copies per ml achieved ≥ 92 % sensitivity, whereas in the standard procedure, samples ≥ 207 RNA copies/ml achieved 100 % sensitivity. Moreover, the COBAS AmpliScreen™ HIV-1 Test, version 1.5 (Roche) is suitable for detecting prevailing HIV-1 variants.

Six-year pilot study on nucleic acid testing for blood donations in China

A six-year pilot study on nucleic acid testing for HBV, HCV and HIV-1 has been undertaken on sero-negative plasmas in mini-pool and individual donation testing at Shenzhen Blood Center. Of 307,740 sero-negative blood samples, 95 of 102 HBV DNA yields were confirmed positive, 80/95 (84.2%) were classified as occult HBV infection (OBI) and 15 (15.8%) as window period cases. Amongst OBIs, 45% carried anti-HBc only, 41.3% anti-HBc and anti-HBs and 13.7% anti-HBs only. HBV DNA yield was 1:3239. One HCV WP and one HIV-1 infected donations were detected. High residual risk was found in current blood donations screening in China.

Hepatitis E virus in Scottish blood donors

Published prevalence figures for hepatitis E virus (HEV) reveal significant regional differences. Several studies have reported virus transmission via blood transfusion. The aim of this study was to establish HEV seroprevalence and investigate a potential HEV RNA presence in Scottish blood donors. IgG and IgM were determined in individual serum samples. HEV RNA was investigated in plasma mini-pools corresponding to 43560 individual donations using nested PCR. Samples amenable to reamplification with primers from a different region were considered confirmed positives, sequenced and analysed. A total of 73 of 1559 tested individual sera (4·7%) were IgG positive, none tested positive for IgM. Plasma mini-pool testing revealed an HEV RNA frequency of 1 in 14520 donations. Three confirmed positives belonged, as expected to genotype 3. HEV IgG and RNA figures in Scottish blood donors are lower than those published for the rest of the UK, but sufficiently high to prompt further studies on potential transmission rates and effects of HEV infection, especially for immunosuppressed individuals.

Hepatitis B virus nucleic acid testing in Chinese blood donors with normal and elevated alanine aminotransferase

Nucleic acid testing (NAT) is currently not a routine donor test in China. The aim of this study was to evaluate the current residual risk of hepatitis B virus (HBV) transmission and the value of ALT testing in preventing HBV infection. From January 2008 to September 2009, a total of 5521 qualified donations by routine screening and 5034 deferred donations due to elevated ALT alone were collected from five blood centers. Samples were tested for HBV DNA by triplex individual-donation (ID)-NAT (ULTRIO assay, on the TIGRIS system, Novartis Diagnostics). HBV NAT-reactive samples were further analyzed by HBV serology, alternative NAT, and viral load and were diluted to simulate if they could be detected in a minipool-NAT. There was no significant difference in the HBV NAT-yield rate between the qualified donations group (5/5521) and the deferred donations group (4/5034). Of these nine potential HBV-yield cases, one donor (11%) was a possible HBV window-period donor, one (11%) was a chronic HBV carrier, and seven (78%) had probable or confirmed occult HBV infections. Of seven potential HBV-yield cases quantified, the viral loads were less than or equal to 70.0 IU/mL. Minipool testing (minipools of 4, 8, and 16 donations) would miss 43% to 79% of the nine HBV-yield donations. Based on our findings in qualified donations, we estimate that the nationwide implementation of ID-NAT testing for HBV DNA in China would detect an additional 9964 viremic donations per year. ALT testing seems to have no significant value in preventing transfusion-transmitted HBV infection. ID-NAT versus simulated minipool-NAT using the ULTRIO test demonstrates the benefit to implement a more sensitive NAT strategy in regions of high HBV endemicity.

Evaluation of pooled rapid HIV antibody screening of patients admitted to a San Diego Hospital

Current HIV screening guidelines in the United States recommend expanding the scope of HIV screening to include routine screening in health care settings; however, this will require increased resources. Since testing of pooled samples can decrease costs, the test characteristics of pooled rapid antibody testing were determined and optimal pool sizes were estimated for populations with HIV prevalence ranging from 0.25% to 10%. Based on these results, pooled testing methods were evaluated for screening patients admitted to hospital in San Diego, California. Evaluation of pooled antibody testing on samples collected from individuals with known HIV infection found only a modest reduction in sensitivity. These false negative results were only found among samples with very low optical density readings (<0.125 by the ADVIA Centaur® HIV assay). These readings are considered as HIV negative by the ADVIA Centaur® HIV assay, and therefore likely correspond to samples collected during acute infection. Further evaluation of pooled testing of samples collected from individuals during recent infection, found that mini-pool testing of five samples detected HIV antibody in 86% of samples taken within 60 days of the initial infection and 92% of samples taken within 90 days of the initial infection. Based on estimations of optimal pool sizes for low prevalence populations, it was decided to evaluate mini-pools consisting of 10 samples to screen the study's hospitalized patients. During this evaluation, the HIV prevalence among hospitalized patients was 0.8%, and the 10 sample mini-pool testing had 100% sensitivity and specificity. Additionally, pooled testing resulted in an 84.5% reduction in the number of rapid HIV antibody tests needed, as compared to testing each sample individually. Even when incorporating the increased costs of technician time, mini-pooled tested would have resulted in a net savings of 8760 USD for the 523 samples tested in the study. Taken together, these results indicate that pooled rapid antibody testing may reduce substantially the costs for HIV screening in low prevalence populations without a loss in accuracy.

A cost–benefit analysis of blood donor vaccination as an alternative to additional DNA testing for reducing transfusion transmission of hepatitis B virus

A survey-based, cost–benefit analysis was performed comparing blood screening strategies with vaccination strategies for the reduction of transfusion transmission of HBV. 231 whole blood donors and 126 apheresis donors were eligible and completed a questionnaire detailing their donation habits. The cost–benefit analysis included current mandatory HBV testing (HbsAg + anti-Hbc, A1), A1 with additional nucleic acid testing (NAT) for minipool (A2) or individual donation testing (A3), as well as HBV vaccination strategies using time-dependant (B1) or titre dependent booster vaccination solely (B2), or B2 in addition to current mandatory testing procedures (B3). Different cost models were applied using a 5% rate of discount. Absolute costs for current mandatory testing procedures (A1) over 20 years in Germany were €1009 million. Additional NAT would lead to incremental costs of 43% (A2) or 339% (A3), respectively. Vaccination strategies B1 and B2 showed cost-reductions relative to A1 of 30% and 14%, respectively. The number of remaining HBV infections could be reduced from 360 (for A1) to 13, using vaccination, compared with 144 or 105 remaining infections for A2 or A3, respectively. Absolute cost per prevented infection is similar (€2.0 million) for A2 and B3. HBV vaccination offers the near-elimination of transfusion infections while representing a potential cost-reduction.

West Nile virus in 2006 and 2007: the Canadian Blood Services' experience

The Canadian blood supply has been screened for West Nile virus (WNV) since 2003. A strategy for targeted individual-donation nucleic acid testing (ID-NAT) was implemented in 2004 to identify potentially infectious donations that may be missed by minipool (MP) testing. In 2007, Canada experienced a larger epidemic than in previous years providing an opportunity to evaluate the ID-NAT triggering algorithm in higher-risk areas. A specially created database and internal-external communication identified regions for targeted ID-NAT using MP and community triggers. WNV-positive donations identified by ID-NAT were reexamined in MP to assess the efficacy of targeted ID-NAT in identifying potentially infectious donations that may have been missed by MP testing. WNV-positive donation data from 2006 and 2007 were analyzed to examine temporal and geographic trends. A telephone survey about symptoms was carried out after the 2007 season. In total 78 WNV-positive donations were identified (66 true-positives and four false-positives being in 2007). Most positive donations were in the late summer, concentrated in the same western provinces as community cases. Fifty-two donations were identified by ID-NAT and 46% were consistently positive in MP. Of the other 54%, 74% were immunoglobulin (Ig)M- and/or IgG-positive. Fifty-six percent of donors experienced mostly mild symptoms before or after donation (but all said they were well at the time of donation). WNV-positive donations correspond geographically with the epidemic. MP testing identifies most potentially infectious donations with a smaller potential benefit from targeted ID-NAT. Mild symptoms are common but may not deter donation.

The efficacy of individual‐donation and minipool testing to detect low‐level hepatitis B virus DNA in Taiwan

Financial constraints are the main concern in implementing nucleic acid testing (NAT) as routine blood screening in Taiwan. The PROCLEIX ULTRIO assay (Ultrio) on the TIGRIS System (Novartis Diagnostics) was evaluated for its operational performance both for individual-donation testing (IDT) and in minipools of 4 (MP4) to develop a feasible solution. Analytical sensitivity was determined by testing WHO international standards. We tested 10,290 blood donors, 4210 in IDT and 6080 in MP4. Potential hepatitis B virus (HBV) yield donors (hepatitis B surface antigen [HBsAg] negative/NAT reactive) were evaluated for up to 9 months' follow-up. Discordant results between the Ultrio assay and the HBsAg tests were further analyzed by HBV antibody serology, alternative NATs, HBV DNA quantification, and sequencing. The 95% limits of detection in IU/mL (95% confidence interval) were as follows: human immunodeficiency virus Type 1 (HIV-1), 18 (12-34); hepatitis C virus (HCV), 4.4 (2.8-8.9); and HBV, 6.3 (4.4-11). The retest rates were 0.55% for IDT and 0.33% for MP4. No HIV or HCV yield cases were found, while there were 12 potential HBV yield cases, nine from IDT and three from MP4 testing. Eleven of them were successfully genotyped as B2. Ten of them returned for follow-up and mostly were determined as occult HBV infection (OBI). The IDT yield rate of 9 in 4210 (0.21%) was fourfold greater than the MP4 yield rate of 3 in 6080 (0.05%; p < 0.05). The higher yield rate for IDT versus MP4 demonstrates the benefit to implement a more sensitive NAT strategy in regions having significant OBI carriers such as Taiwan.

West Nile virus testing experience in 2007: evaluation of different criteria for triggering individual‐donation nucleic acid testing

In 2007, clients served by Blood Systems Laboratories used variable approaches for triggering West Nile virus (WNV) RNA individual-donation (ID) nucleic acid testing (NAT). These included two minipool (MP) NAT-reactive donations and a greater than 1:1000 rate in a 7-day interval (primary trigger), criteria based on one MP-NAT-reactive donation when there was WNV activity in overlapping and/or adjacent geographic areas (neighbor trigger), or zero MP-NAT-reactive donation (self-trigger). The Procleix WNV assay was used in either a 16-sample MP or an ID format. NAT-repeat reactivity or anti-immunoglobulin M (IgM) positivity defined true positives (TPs). TPs that were negative on 1:16 dilution testing were considered ID-NAT yield cases. WNV NAT performed on 1,217,929 donations identified 162 TPs; 87 were detected by MP (rate of 0.008%) and 75 by ID (rate of 0.10%; p < 0.0001). There were 34 ID-NAT yield cases, including 4 IgM/immunoglobulin G (IgG)-negative and 9 IgM-positive/IgG-negative donations. Rates of yield cases by primary, neighbor, and self-triggering were 0.077, 0.052, and 0.004% (p = 0.0003). None of 11 ID-NAT yield cases detected by the neighbor trigger would have been detected if the primary trigger had been used. Primary triggering criteria identified 21 viremic donations that would have been missed by MP testing; however, 11 other low-level viremic donations required more stringent criteria (e.g., neighbor trigger) for detection. It is reasonable to adopt more stringent ID-NAT triggers, including elimination of the rate criterion and triggering on one NAT-reactive donation for regions adjacent to centers which have already triggered.

Hepatitis B virus traceback and lookback: factors to consider

Hepatitis B virus traceback and lookback: factors to consider

Performance characteristics of the Food and Drug Administration–licensed Roche Cobas TaqScreen West Nile virus assay

The cobas TaqScreen West Nile virus (WNV) test (Roche Molecular Systems) was licensed by the Food and Drug Administration (FDA) in August 2007 for detecting WNV RNA in pools of six or in individual donations (IDs). A series of studies established the performance characteristics of the assay and test system before FDA licensure. Analytic sensitivity was determined by probit analysis using multiple source materials. Clinical sensitivity was determined by testing a panel of 315 known WNV RNA-positive specimens. A large clinical specificity study was conducted by five laboratories during months when WNV activity was not expected. The 95 percent limit of detection for ID testing using the Lineage 1 Health Canada WNV reference standard was 40.3 copies per mL (95% individual donation, 35.1-47.8 copies per mL). Clinical sensitivity was 100 percent (95% confidence interval [CI], 98.8%-100%) for ID testing and 97.5 percent (95% CI, 95.1-98.9%) for minipool (MP) testing. Clinical specificity, when resolved to the ID, was 100 percent for both formats and was 99.986 percent at the MP level. The cobas TaqScreen WNV test performed on the cobas s 201 system is a fully automated test system with excellent clinical sensitivity and specificity that offers the benefits of automated sample preparation and a secure environment for donor testing information.

Supply with Platelet Concentrates from the Point of View of a Blood Donation Service of the Bavarian Red Cross

Background: It is known that pooled platelet concentrates derived from buffy coat (PCs) have several disadvantages compared to platelet concentrates produced by platelet apheresis (APCs). Therefore, all blood products issued by the Bavarian Red Cross blood banks (BSD/BRK) (18,000 products/year) were produced by single donor apheresis. The main reason not to produce PCs was the elevated viral and bacterial infection risk during the last decade. But also the four-fold increased exposition to HLA and PLA antigens and the poor quality (in the sense of white and red cell contamination) of PCs (especially the ones produced with the platelet-rich plasma method) played a role to abstain from these products. Material and Methods: We performed a risk assessment to evaluate both products with regard to the actual testing and production methods, considering recently published data. However, a statistical calculation of the risks associated with the use of PCs or APCs with regard to different infectious agents with various prevalences was not done. Results: The dramatically reduced risk for the transmission of HIV, HBV or HCV accompanying the implementation of improved antibody tests and of NAT minipool testing, the introduction of 100% leukocyte filtration, the conversion of PC production from the platelet-rich to the buffy coat method, and recent data on the risk of transmission of bacterial infections resulted in a equal assessment of APCs and PCs. Conclusion: As a consequence of this revised risk assessment, we supply our hospitals with both products APCs and buffy coat-derived PCs (pools of 4 donors). For clinical use we considered both products as equally effective, except for patients who have multiple antibodies and need HLA-typed platelets.

Testing Strategy To Identify Cases of Acute Hepatitis C Virus (HCV) Infection and To Project HCV Incidence Rates

Surveillance for hepatitis C virus (HCV) is limited by the challenge of differentiating between acute and chronic infections. In this study, we evaluate a cross-sectional testing strategy that identifies individuals with acute HCV infection and we estimate HCV incidence. Anti-HCV-negative persons from four populations with various risks, i.e., blood donors, Veterans Administration (VA) patients, young injection drug users (IDU), and older IDU, were screened for HCV RNA by minipool or individual sample nucleic acid testing (NAT). The number of detected viremic seronegative infections was combined with the duration of the preseroconversion NAT-positive window period (derived from analysis of frequent serial samples from plasma donors followed from NAT detection to seroconversion) to estimate annual HCV incidence rates. Projected incidence rates were compared to observed incidence rates. Projected HCV incidence rates per 100 person-years were 0.0042 (95% confidence interval [95% CI], 0.0025 to 0.007) for blood donors, 0.86 (95% CI, 0.02 to 0.71) for VA patients, 39.8 (95% CI, 25.9 to 53.7) for young IDU, and 53.7 (95% CI, 23.4 to 108.8) for older IDU. Projected rates were most similar to observed incidence rates for young IDU (33.4; 95% CI, 28.0 to 39.9). This study demonstrates the value of applying a cross-sectional screening strategy to detect acute HCV infections and to estimate HCV incidence.

Balance module allows consistent monitoring and documentation of the pooling process for nucleic acid amplification technology testing

When performing minipool testing of blood donations for transfusion-transmissible viruses, it is crucial to correctly dispense plasma from each donation into the pools. However, concerns regarding the monitoring and documentation of the pooling process exist. A balance module with a tube holder has been developed, which can be easily integrated into liquid handling platforms. The existing software monitors and evaluates every single dispensing from primary samples to the minipool to confirm liquid arrival. The weighing accuracy of the balance module is approximately 2 mg per dispensing episode. Ten thousand one hundred and fifty-six blood donations were tested on a Tecan Genesis RSP pipetting system. Thirty-one donations were not pipetted because the pipetting workstation identified a clot or sample shortage in the primary tube. The balance module exclusively detected another 18 mispipettings, which were outside of the acceptance criteria of 100 +/- 10%. The mean pipetted volume for these samples was 34.2 microl (range 0-87 microl). Visual inspection of the corresponding primary tubes showed blood clots, short sample or no apparent cause. The average deceleration of the pooling process, using the balance, was determined to be about 22%. With the novel liquid arrival check system, complete and consistent process documentation of pooling for nucleic acid amplification technology (NAT) testing is feasible. It enables blood banks to monitor and compare every single dispense made with predefined, required volumes. Results can be transferred to the laboratory information management system for automated selective exclusion of inaccurately dispensed samples. ONE SENTENCE SUMMARY: Monitoring and documenting the pooling process for NAT testing using a balance-based liquid arrival check system.