diff --git a/testsuite/MDAnalysisTests/analysis/test_base.py b/testsuite/MDAnalysisTests/analysis/test_base.py
index 90887b2ad0..e2fe428376 100644
--- a/testsuite/MDAnalysisTests/analysis/test_base.py
+++ b/testsuite/MDAnalysisTests/analysis/test_base.py
@@ -121,6 +121,17 @@ def test_incompatible_n_workers(u):
     with pytest.raises(ValueError):
         FrameAnalysis(u).run(backend=backend, n_workers=3)
 
+
+def test_n_workers_conflict_raises_value_error(u):
+    backend_instance = ManyWorkersBackend(n_workers=4)
+
+    with pytest.raises(ValueError, match="n_workers specified twice"):
+        FrameAnalysis(u.trajectory).run(
+            backend=backend_instance,
+            n_workers=1,
+            unsupported_backend=True
+        )
+
 @pytest.mark.parametrize('run_class,backend,n_workers', [
     (Parallelizable, 'not-existing-backend', 2),
     (Parallelizable, 'not-existing-backend', None),